Tipping point Essay Example | Topics and Well Written Essays - 750 words

Tipping point - Essay Example joinings are perceived to be the trendsetters because of their rich network of acquaintances and friends. They know many multitude and keep in touch with them. Simply put, connectors strive to bring the world together not for their benefit, but for the benefit of humanity. On the other hand, Mavens are populate who take delight in finding new information and share that information with connectors. Unlike connectors who just wish to move over a vast network of friends around them, mavens strive to have hottest esoteric scoops on specials and gadgets, and share that information with consumers. Hence, they fundamentally connect consumers to the marketplace due to their aptitude and capability to be informed on functionality of products, prices, and market trends. While the Mavens provide the message and connectors spread it, salesmen are skilled in persuading people to believe in what they hear. Their ability to easily build a rapport makes them good in expressing feelings and emotions, and very much use their charisma to manipulate others. A striking difference between salesmen and connectors is the sheer fact that connectors depend not as individuals, but in nodes and are to a greater extent obliged to first help you then(prenominal) money later however, salesmen are apparently money-oriented as they hugely motivated by money when box and pricing innovations (Gladwell, 82). Similarly, while connectors seem to rely on more general information to connect the world, Mavens gibe deep into ideas and innovations and give detailed and unambiguous information regarding it.Apparently, I am more similar to the Connector personality type. Over the past few decades, with the dawn of technology I have been finding myself more engaged with the social media including Facebook and Twitter. I start each day on Facebook and Twitter by bankers bill and twitting unpretentious questions such as How can I help you?, Is anything the matter? Th rough these platforms, people have been able to express

Human Sexuality Aspects Essay Example | Topics and Well Written Essays - 500 words

Human Sexuality Aspects - Essay ExampleHuman sexuality deals more with emotional responses as principles, standards, and duties. Movies as a media of communication, has an enormous effect in the manner in which it has shaped the way population relate and engage in sexual activities. Human sexuality directly or indirectly has influences in many areas of life that pertains to human beings.Human sexuality affects the society in numerous ways. For example, it influences persons in the sociocultural aspect. Different people have different cultures and share different views on sexual behaviors and what people perceives to be wrong in different parts of the world. Movies have played a played a major role in influencing the way people perceive sexual activities. For instance, most characters in movies engage in sexual activities before marriage, which is a forbidden practice among many religious groups. Though most persons allow for try their best to follow religious rules, they will also try their very best to hap secrets of how they engage in sexual activities. People do this in order to avoid critisims or change surface being punished. Moreover, sexual activities in movies in addition to beliefs of a person and their religious backgrounds as swell as personal experiences and behaviors can have even a stronger influence in an individual or a group of people. Such influences from movies include influencing the way children are taught about sexuality by their parents. This is because children turn tail to emulate in behavior what they watch on movies. Often, young children look for guidance through media on what are both socially acceptable or not. The media tends to help shape the attitudes of children on what they like and what they do not. Movies show attractive cast people in the lead roles (Bancroft 7).Human sexuality also plays a major role in influencing the way people in the society behaves and communicates to each other. More very much than not, human behavior is determined by the

Management Information systems - Case study Essay

Management Information clays - Case study - Essay ExampleAdditionally, the Maintenix system is available on the Web as well as easy to utilize and could be complete for working and operable stations all through the world. In addition, it lessens cyclic tasks and protracted imperfect or missing info and culture by permitting engineering, maintenance and finance divisions to simply share corporate data and information. Moreover, wireless technology based arrangement makes Maintenix information system more efficient like equipment, atmosphere technicians and parts are for all time on the move. Matintenix offered very effective system arrangement that is complaisant on the Web and simple in managing all the stations all through the business domains, regarding operational and functional competence. In addition, software Matintenix minimizes repetitive jobs and missing time tracking or imperfect information by permitting engineering, maintenance and finance divisions to simply share data and information across the globe. Maintenix software employs a contemporary and up-to-date architecture and offers superior potential like that function-based computerized workflow, Web browser interface, electronic database signatures, superfluous adapters and facility for portable wireless devices.

Wilma Mankiller video Essay Example | Topics and Well Written Essays - 250 words

Wilma Mankiller video - Essay vitrineFor instance, she explains that when people want to know about the current issues facing the internal Americans, they should first understand the past. She adds on this by stating that it takes the enduring strength of the Native Americans to go through the history they have had.ManKiller continues to give begin on issues she feels the U.S. public should understand in totality. In her consistent embrace for the tribal ways, she explains that the tribal governing body prefers to take care of some issues. These include foster care, adoptions and privileges for water rights (Mankiller 100). Through this project, they have managed to avail wellness facilities to the locals. Clearly, this is one significant point that cannot be missed from the speech. In a way, she embraces the changes happening in the Native American societies, but mainly insists that it is necessary to keep the cultures. Interestingly, she bounces the speech with bits of sense of humor about a tourist visiting the Cherokee nation hoping to find the native Indians in full attire. However, to the tourists disappointment, Theyre probably all at K-mart. (Michael

Bus 234 Quail P1 Essay Example | Topics and Well Written Essays - 250 words

mickle 234 Quail P1 - Essay ExampleAll field agents have an educational background in environmental science while the accountant has an educational background in accounting. The mission of the business is to provide forest services in terms of advising clients on the best approach to designing and putting up foot to allow minimization of environmental pollution in order to make the immediate surrounding fitter and friendlier.The business anticipates to add the number of clients through the field agents. The field agents interact with clients on site and whitethorn attract new clients from their code of conduct and interaction with existing clients on the ground. The company wishes to conduct a needs assessment to determine the necessity of training its field agents on the best strategies for winning potential customers to utilize the business services. As discussed in Altschuld and David (57), in order to determine the training needs, all the field agents were given questionnair es, which contained open-ended questions on a Likert scale.From the results obtained from the short survey, the field agents rate of interaction with prospective clients is median(a) since their interaction skills and convincing are also rated average. It is thus imperative for the Earthcare group to train its field agents on best skills and strategies of developing and sustaining good customer relationships in order to increase the customer base for Earthcare

Juvenile probation etc Essay Example | Topics and Well Written Essays - 1500 words

Juvenile probation etc - Essay ExampleAccording to the chief components present in the IAP model, the political program greatly aims at diminishing recidivism through intensive supervision and control measures, strengthening family and biotic community relations with individualized case planning and reinforcing progress in the improvement of personal well-being by incentives and an enthusiastic case management utilizing resources within the community (Wiebush, McNulty, & Le, 2000). Failures however in meeting certain objectives of IAP return from implementation problems such as staffing vacancies, institutional crowding, and unstable program environments. Successful operation nevertheless takes into write up the consistent adherence to the core characteristics of the model and that it is found to establish more strength for the institution in collaborating with the external support teams of afterc atomic number 18 and research. increment, on the other hand, specific altogethery provides intensive aftercare for high-risk female juvenile offenders. Managed by Boys and Girls Clubs of South Alabama, Inc. with the help of the Office of Juvenile Justice and Delinquency Prevention, this program makes use of Network Aftercare body (NAS), an IAP adaptation. Principal components are inclusive of female-specific life skills, adventure therapy, community service, academic education, and Functional Family Therapy. Female offenders are administered a one-year aftercare monitoring and supervision upon release from detention. GROWTH keeps up with the similar goal to take recidivism as well as pregnancy rates and to increase job placements for GROWTH dispelicipants and these are all part of the programs documented success. No significant failure has been evaluated to emerge during program implementation as GROWTH promotes additional services such as individual and group therapy, substance abuse treatment, and mentoring. Multidimensional intervention Foster Care (MTFC) being another aid in a stage after probation, create at the Oregon Social Learning piazza, provides highly trained foster parents, intensive support from a clinical team, and a tightly structured program. Besides becoming less prone to slip back up into old delinquent patterns of behavior, MTFC sets goal that ensures better peer associations and development of both academic and professional skills. The program gives sufficient focus on individualized treatment where behavior is monitored and recorded on a day-to-day basis through contacting the foster parents via the telephone. No ample amount of data shows evidence of the programs shortcomings and as part of the programs success, MTFC participants experience fewer arrests, minimal involvement or interest in delinquent activities and lesser timeframe of incarceration. Facilitating males of ages 13 to 18 for whom the Maryland Department of Juvenile Services render committed obligations, Thomas OFarrell Youth Center serves treatme nt concentrated on the development of positive social norms. Planning for aftercare begins at admission to residential treatment where youth spend an average of eight months. While in residential care, youths are appoint two caseworkers each and these persons are required to look

Health Program For Latina Immigrant Essay Example | Topics and Well Written Essays - 750 words

Health Program For Latina Immigrant - Essay ExampleThe other some important thing that I should be aware is the mode of greeting in which virtually of the Latin people are welcoming and mostly use hugs and cheek kisses as a mode of greeting. shrewd this impart ensure I behave according to in order to show my gratitude. Politeness is similarly a key value and thus noting that interrupting someone shows disrespect. I leave huddle this during the class period. pietism and family are highly valued and thus being conversant with the Roman Catholic Church doctrines will make it easier not to respect them in my teachings. Most of the Latin Immigrants are not well up better thus using a program in Spanish to suit them, this is better than translating a program made in English. The props used should be also in Spanish and as an educator I would involve Spanish entertainment and their cultural foods to win their hearts. Since most of them are not well educated, it is wise to ESL app roach, which will captivate more interest with all the people that want to learn English. Lastly, I would participate fully with the church thus create an admirable image thus people would be welcoming and willing to listen to me.Would your program be for the person, extended family, or neighborhood?My program will be for the extended family since the Latin culture values it most. This will make my bonding experience easier since most of the immigrants have no documents and are afraid of being deported. These immigrants will secure to have classes in their homes.

To what extent was the involvement of Winston Churchill during the Essay

To what extent was the involvement of Winston Churchill during the violation of Gallipoli ultimately responsible for its harm - Essay Exampleobsolete and not much use for a battle against the High Seas Fleet, the operation would make no difference to the numerical superiority of the Royal Navy. The failure of its execution nearly destroyed his political career, and tarnished his reputation in the eyes of many contemporaries.For Great Britain, June 1940 was round into one of the worst months of its modern history. After the swift fall of Belgium, Holland and France via the German Blitzkrieg, the British found themselves alone alone in confronting what seemed to be the invincible state of war machine of the Third Reich. Many asked themselves at this prison term whether Winston Churchill, who had only been Prime Minister for a few weeks, was the right person to lead the nation at such a difficult time. In his favour he had his unbreakable faith in success and unmatched political experience. Nevertheless, there were those who remembered his stubbornness and adventurous character during the First World War, and were convinced that he had led the United Kingdom to spectacular failure at Gallipoli but was Churchill truly to blame for that wipe out?In the Autumn of 1914, barely three months after the beginning of the First World War, the land war had reached a stalemate on the Western Front.1 On the Eastern Front there were signs that the Germans would eventually have the best Russia, especially after Turkey blocked off supplies from Britain and France. After the First Battles of the Marne and Ypres, the Schlieffen Plan, which was supposed to open the route to Paris in the same way that it had been achieved in 1870, had failed.2 In its place there was the beginning of a war of attrition which promised to be long and difficult, and which used up all the resources of the combatants. Its symbol was to be deep warfare in which thousands of men would die for each disputed inch of ground. Bloody attrition in which the battles would end with hardly any change to the position of the Front. The various High Commands

Tesco Personal Finance Case Study Example | Topics and Well Written Essays - 4000 words

Tesco own(prenominal) Finance - Case Study ExampleThis encyclopaedism of Tesco over the 50% stake of RBS coming as a direct challenge for the high street monetary institutions which currently are on the inclement side of the economic recession tide and registerly are in-waiting for the bank charges decision by the spicy Court.The acquisition will help Tesco to make its involution from financial serve that revolved around an miscellanea of popular financial services and products to full-fledged retail banking oblation more preference, innovation and value to customers and repair returns to its shareholders. It is a part of Tescos strategy through which Tesco will be able to further enhance its festering in the services market sector that includes telecoms and internet/home shopping as well. According to Tescos theme for the year 2008, by taking the full ownership of the TPF, Tesco would be able to increase its annual moolah to 1 billion from 240 million (News Release, 20 08).This acquisition clearly reflects that Tesco has identified considerable scope for further affirmable developments and advancements in the financial sector which is itself a very extensive and a dynamic sector. Hence by taking the full ownership, Tesco will be better equipped to be able to exploit the prospective offered by the TPF in the more basic version of banking. This second half of the paper examines the strengths and weaknesses of Tesco hence offering an analysation of the resources and capabilities that Tesco can muster up to compete effectively and develop TPF into a full service retail bank making a success of full- fledged consumer banking. Tescos Expansion to Full Service Consumer BankingOn the acquisition of RBS stake and TPFs expansion into full- fledged retail banking in the coming years, Sir Ted Leahy, the COE of Tesco plc reflected, As consumers look to make both pound work harder it is a good era for Tesco to expand its presence (BBC, 2008). The phrase good time refers to the current financial turmoil, which in recent times has revealed the shaky foundations of many a high compose financial institution, and ironically has presented an opportunity for TPF to move into full service banking. As a result of the financial depression many of the TPFs would be competitors will be at their weakest, hence competition that will be on offer will also be comparatively less.In order to make the expansion into the more core-sphere of banking, the management of Tesco will need to rally up their strengths comprising its assets and resources identify with the current financial trends the opportunities or threats presented by it and also realise their own weaknesses and accordingly create business objectives and strategies which will allow the best use of resources and strengths, maximum exploitation of opportunities, stablisation of threats and minimisation of weaknesses and other draw backs for the overall success Tescos expansion endeavor.For the present short time frame, Tescos strategic objective for the future development of TPF is to broaden the horizons of the existing financial services rendered by TPF, which includes services and products like savings accounts, credit card accounts and insurances and moreover increase its presence in the Tesco stores. However, on

Mutual Fund Regulation Dissertation Example | Topics and Well Written Essays - 6250 words

shared Fund order - Dissertation ExampleWe affirm the lower courts decision. Background The case records reveal that on March 31, 2011, the US regularise Court for the Southern District of New York denied a class work related to the fall in mutual fund share prices during the 2008 global financial crisis. The plaintiffs claims were made consistent(predicate) to components 11 and 12(1)(2) of the Securities shape of 1933 (hereinafter the Securities Act). Specifically, the plaintiffs claimed that the relevant prospectus issued by the SSgA Yield Plus Fund misrepresented the Mutual Funds risk exposure to mortgage associated securities. The US District Court for the Southern District of New York denied the plaintiffs claims on the grounds of loss origin pursuant to Section 10 (b) of the Securities Act. It was held that under Sections 11 and 12(a)(2) of the Securities Act, the allowable quantification of damages is confined to a decrease in the value of securities that follows from the transmission of fabricated swelling/misrepresentation of the purchase price of security. Since mutual fund shares may only be sold or redeemed at a statutorily formulated price based on the net summation value (NAV) of the funds securities, misrepresentations in the corresponding prospectus cannot inflate the NAV of the mutual fund and gum olibanum cause a fall in the NAV. Therefore the lower court held that the plaintiffs charge did not properly plead the required loss causation and the action was subsequently dismissed with evil (Yu v. State Corporation, 2011). The appellate claimed that the document offering the mutual funds misrepresented the nature of the securities or enthronization held by the Yield Plus Fund, misrepresented its description, the Funds objectives and the risks associated with mortgage exposures and the risk associated with place in the Fund (Yu v State track Corporation, 2011). The Respondent claims that the Appellants motion should be dismissed b ecause (as previously argued) the Appellant did not plead falsity and materiality (Yu v State Street Corporation, 2011). The Respondent also argue once again that the certifications that they indorsed did not contain false statements of material facts the Appellants claims should be denied because the Respondents are not vendors of the mutual funds and the control person claims under Section 15 should be dismissed (Yu v State Street Corporation, 2011). The Respondents also claim that the loss causation is not substantiated because even if the statements had been substantiated, the damages claimed are not linked to the alleged misrepresentations or omissions relied on. Opinion Securities Act of 1933, Sections 11(a) and 12 (a)(2) When investors purchase mutual fund shares based on a mutual funds published statement and the latter reflects a material misstatement, investors have wide remedial recourse pursuant to Sections 11(a) and 12 (a)(2) of the Securities Act. The investors are abl e to recover the resulting decline in the value of the shares without having to claim reliance upon the misstatement or without having to prove that the defendant was somehow culpable in terms of the misstatement. Additionally, the claims under Section 11 (a) and 12(1)(2) do not call upon the shareholders to comply with

The essay should be presented as a Journal opinion article. The

The should be presented as a Journal opinion condition. The article should be timely and address an stinting bulge out of curren - Essay ExampleThe brusk and abundant term implications of immigration laws be imperative for determining piece of work rates and productivity of the US workforce. In the short discharge, immigration is said to slightly decrease the appointment rates or in other words, increase unemployment rates however the long run implications are ordained. The matter has been the center of debate and has affected the US immigration policy greatly. While many translate to make a case against reform in US immigration laws, credible sources in the acquirable literature have established that long term bear upons of immigration on the US workforce is positive as it enhances labor productivity. In their opinion article titled more(prenominal) Immigration Means More Jobs for Americans, John Dearie and Courtney Geduldig have given useful statistics that suggest tha t immigration improves employment as it creates jobs for the mutual people (n.pag.). It mentions how immigrants make up on a small percent of the total American universe of discourse yet contribute a significant percent to the economy by means of small businesses. Because intimately immigrants are self-employed and own several small scale sole-proprietorships and partnership businesses, they create jobs for the people thereby service to reducing the unemployment rates. ... ed by Dearie and Geduldig is consistent with current findings which Giovanni Peri has described in his article The Impact of Immigrants in Recession and Economic Expansion. Peris article mentions the long run positive effects of immigration on the US economy as it increases productivity (4). Immigrants bring in new ideas and creative thinking along with them leading diverse distribution of organizational workforces. Although it may seem like immigrations negatively impact the US labor market as they saturate jobs and cause unemployment in the short run, the long run implications are promising. However, it seems to long before the long run positive impacts become observable. Even though short run effects of immigration may reduce employment, it increases average income in the long run (Peri, 4). So, the long effects of immigration are positive on employment, productivity, and income. However, it is also important to note that the immigration moldiness adapt to economic changes in station for it to improve variables such as employment, productivity, and average income. The current US immigration laws do not allow the immigrants to adjust to economic changes because it makes them unresponsive to the economic conditions. In order for legal immigration to improve organizational productivity, average income, and employment rates, immigration must adjust to economic cycles thereby leading to improvements in the US economy. These adjustments are the hardest to carry during prolonged periods of economic downturns and therefore amendments in the US immigration policy is required to allow the inflows to be responsive to economic changes thereby helping to boost economic activity and hence result in greater jobs for the people. Thus, a reform in the current US immigration laws may provide a

A survey on knowledge, attitudes and behaviors of young adults about Essay

A subject field on knowledge, attitudes and behaviors of young adults about health promotion and heart disease - Essay ExampleThe playing field can also include the Preventive Medicine Attitudes and Activities Questionnaire (PMAAQ).Survey type research studies usually scram coarser samples because the percentage of responses generally happens to be low. Thus, the survey method gathers data from a relatively large number of cases at a particular time. The degree of precision increases if the sample size is larger and decreases if the sample size is smaller. Power analysis helps to determine the sample size. About 500 subjects can be include for this study.Purpose of this research is to explore the young adults knowledge, attitudes and behavior about health promotion and heart disease. In health and human development, young adulthood is considered to be the stage between adolescence and adulthood, roughly ages 16 to 30. Literatures show up that generally young adults had a poor d egree of knowledge and poor health behavior link up to heart disease.The study participants will be administered with a structured questionnaire. Otherwise, the questionnaire can be mailed to the respondents with a request to return after completing the same. The participants are informed to answer all the questions.

The Conglomerates, the Lovers, the Spies and the Outcast Essay

The Conglomerates, the Lovers, the Spies and the Outcast - Essay ExampleThe conglomerates tog parties, treats and keep looking for occasions of celebration. They atomic number 18 life long friends. They whitethorn go to different places subsequently school, yet call one another daily. Mostly, they land in the alike(p) profession and in the same country.The lovers are groups of two. A third one is not allowed. The two are inseparable. Their academic performance may differ significantly. Lovers may have different interests, but they are attached emotionally. Many lovers are altogether opposite to each other in nature, yet they make a perfect match. The effect is magnetic, rightful(prenominal) like north and south poles of the magnet attract each other. The lovers remain together full measure at the school and frequently meet each other outside as well. In cast jokes, the lovers are referred to as gays, but the lovers dont care. However, their future is quite unexpected. Usually , people expect lovers to be life-long friends, just like conglomerates, but it hardly ever happens. Most lovers depart after the school is over, neer to see each other again.The spies are individuals. They have no association with any group, but their favorable s cut downs are quite enhanced. They survive individually. Spies have good oratory skills, so make excellent irregular friends. They have no permanent friend. One day, they can be seen with one person, the next day with another. They claim a temporary company. People suspect the spies whenever their secrets leak out. They are generally the most untrustworthy and at the same time, they are the most interesting class-mates around. The spies are witty and cunning.The outcast is also individuals, but with genuinely poor social skills. They are not allowed entry into any group. They are hated and mocked. The outcast are objects of fun for others. The outcast fear the class-mates and hardly ever raise their voice against the injustice they are offered. They kill the time sitting around a corner in the classroom. Surprisingly, many outcast class-mates are exceptionally good in studies, perhaps because they seek refuge in books. The outcast have the most muffled personalities and cant handle mockery.

The Marketing Plan of Crocs Shoes Assignment Example | Topics and Well Written Essays - 750 words

The merchandise Plan of Crocs Shoes - Assignment ExampleThe ac caller is also planning to penetrate in the live markets to increase its sales.The alliance has been growing at a rapid pace since 2002. The company has expanded its trading operations to 125 countries (Cross b). With the increasing sales of the company, the company has been successful in increasing the market share. The sales and gainfulness of the company are presented in the figure belowRecession influenced the sales of the company as the company had shown a significant increase in the revenues and profits from 2006 to 2007. However, from 2007 to 2009, the sales of the company declined. From 2009 and onwards, the sales and profitability of the company have been increasing.The company offers casual footwear for people of all ages including women, men, and children. By now, the company has sold much than 200 million pairs of footwear (Cross a). Some of the major competitors of Crocs are Deckers Outdoor Corp. and S teven Madden (Sharma). The following take in presents the revenues of the three companiesThe company has been expanding its operations globally. To capitalize on the opportunities in the global market, the company has been using diverse strategies including joint venture and acquisition. For instance, the company acquired Jibbitz, Ocean Minded, Tidal Trade, Fury, and others to expand its operations.It is important for Crocs to improve the brand awareness. It has been determine that the company is relatively new in the market and many potential customers do not acknowledge the offerings and quality of the products of the company.Promotion would be crucial to the success of the company in the next few years. The company needs to increase the brand awareness and portray a better brand image in the eyes of the consumers. Therefore the role of promotion will be significant.

Market plan Essay Example | Topics and Well Written Essays - 1250 words

Market plan - see ExampleThus, all in all, the Masafi Group has come up with a winner in the force of the Masafi Natural Mineral Water.The Masafi Water Company is now trying to launch ABC aftershave company, for which they need to practise a step by step foodstuffing plan, since it would assist them in knowing the need of this product, and tax it with respect to the environment in which the industry is thriving at this point in time. (Kerin, 2004)A market plan is basically an outline of the various environmental factors as well as the selling dynamics that will be associated with the product, which is yet to be launched. (Larsen, 1997) It helps the organization to get an aerial view of the viability of the product persuasion that they have, with respect to the company itself, the target customers, the competitors present, the collaborators and the climate. The SWOT analysis also plays a vital role in telling the company about its strengths, weaknesses, opportunities and threat s. The market plan becomes further refined when portions for market segmentation, alternative marketing strategies and selected marketing strategy are added. (Luther, 2001)The Masafi group now plans to launch an aftershave company. Aftershaves are a health and sweetie product and come under the sub-category of fragrances and perfumes. The aftershave market is quite saturated with many big perfume stigmas doing brand extensions into aftershaves, using their known and established brand names. Whether or not Masafi water should plan to invest their time, resources and capability in launching an aftershave company would be better decided if a marketing plan is made, and critically analyzed. (Stapleton, 1998) The marketing plan would consist of the following componentsThe SWOT analysis involves the analysis of the internal strengths and weaknesses of the company as well as the external opportunities and threats that the company is

Delivering Business Value with It at Hefty Hardware Essay Example for Free

Delivering Business Value with It at unchewable Hardware EssayIT Value is defined as the worth or desirability of a thing. It is a subjective assessment. The honor of IT depends very much on how a business and its individual managers choose to soak up it. Strategic positioning, increased productivity, improved decision making, cost savings, or improved service are each(prenominal) ways value could be defined. It is tied to the agreements business model because adding value with IT should alter a firm to do it business better. One company sees value resulting from all parts of the organization having the same processes another defines value by return on investment (ROI) still another measures t by a composite of key performance indicators. IT value proposition is that everyone involved in an IT beginning(a) agree on what value they are trying to deliver and how they will recognized it. IT Value is a function of people, process, and technology.The Three Components of the IT Value Proposition Identification- Recognize and evaluate opportunities. Conversion- Transformation of ideas and opportunities into IT value propositions and creating IT value. Realization- Deliver value and ensure that technology is widely accepted Best Practices in Identifying Potential Value Joint business-IT structure. differentiate value across projects. Utilize a portfolio approach to project selection/prioritization. Establish a patronage mechanism for infrastructure. Conversion Managerial issues with Conversion More projects than resources Insufficient time to complete all projects brusque technical/business resources significant business process redesign may be required for carrying into action of IT.IT value creation can be successfully made by Excellent project circumspection Effective execution (e.g. change management) Reliable IT operations Best Practices in Realizing Value Measure outcomes against judge results.Does Expected Value = Actual Value? Plan a value-rea lization phase for all IT projects Look for and fade root causes of problems Assess value realization at all levels in the organization Have planning for acting on new opportunities.

The Aztecs simplistic crop-enhancing Essay Example for Free

The Aztecs simplistic crop-enhancing studyThe main supply for food for the Aztecs were agriculture and farming. They rely mainly on the crops for every season they grow, induce and consume food and this is how they get their daily food. Terracing Indicating the societal complexity of the Aztecs, the farming technique known as terracing is complicated and elaborate. They used terracing in the hilly areas and built walls, forming terraces into the sides of the hills. These stone walls ran parallel to the contour of the slope and rain washed compost vegetation andnutrients from the hills above. Terracing opened up previously unusable land for farming. Irrigation Because the Aztecs built protracted canal and water-diverting systems, irrigation farming became popular and productive. With water readily available, the Aztecs could enhance the growing season by irrigating the field prior to planting, strengthen their crops by augmenting the rainfall with additional watering, and leng then the farming season by irrigating colossal after the rains had stopped. ChinampasAlso know to the Aztecs as floating gardens, chinampas were pieces of land created by piling up alter layers of compost vegetation and mud from the lake bottom, in the shallow marshy parts of the lakes. Initially the 30 feet by 330 feet chinampas were held in place by man-made posts, precisely eventually these fertile farming patches stayed secured by the grow of nearby willow trees. Once the mud and vegetation piled up higher than the water level, the plot was build for cultivation.The crop yield from chinampas was so prolific and expedient, farmers began growing, not only for themselves, but for marketing out to others. rainfall Cultivation The Aztecs most basic and simplistic crop-enhancing technique, called rainfall cultivation, involved fallowing the fields. They left farming plots empty to collar itself and become nutritionally enhanced by rainfall. This process is the least labor inten sive form of farming, but the Aztecs eventually cast it aside following the population boom and the need for larger, more productive crops.

Geography of the Big Mac Essay Example for Free

Geography of the Big macintosh EssayMcDonalds has been just about since 1940, when it was created by Nick and Mac McDonald in Bernardino, California. Since then McDonalds has only grown around the world in popularity and business. on that point be shortly much than 33 thousand restaurants around the world in 119 countries. The chain has remarkably gone motley offering just a few items on its menu to a wide range of everyplace a 145 diverse items on its menu. Need little(prenominal) to say McDonalds has embedded itself within the worlds society. The way McDonalds runs its business has many different components. These different items include geography of a location, Webers model, victimisation, and mass consumption. One of McDonalds most successful items on its menu is the monstrous mac. Worldwide it is cognise for its flavor however, many do not understand how that burger gets to them. The ingredients ar not merely picked up at a local store, but shipped across miles of land and sea. There be many different occurrenceors for why this happens. The three primary(prenominal) reasons for why this occurs ar due to geography, webers model, and development.Geography plays a boastfully role in where things grow for instance if one looks at the join States corn grows better in the Midwest. This then affects what farmers provide grow in a certain climate, because they need to meet the needs of the bring they atomic number 18 growing. This pattern is seen in Europe with the ingredients needed to influence a big mac and more much than not affects where certain places receive certain items. The buns used to create a big mac are stupefyd in Germany, America, and Canada. The climate in Germany is a mode dictate climate with a good amount of pelting throughout the year.This makes it an ideal place to grow wheat which needs the climate to be mild and not too cold. There are besides many places throughout the United States and Canada that classify as this climate. The pickles are needed to be raised in a warm atmosphere. This is why the pickles for the big mac are grown in Spain and Turkey. Spain has a Mediterranean climate with dry summers and relatively warm winters, and Turkey has bouncy and dry summers and mild winters. The beef is nurtured in Ireland. This is mainly due to the fact that Ireland has sporadic weather that is not necessarily spectacular for growing crops.This means that they are not losing money by not plant a crop but using the land instead is used for cattle ranching. The dairy farmers often need a cool climate to run their farms. This correlates with the fact that Germany and Ireland are the suppliers for the big macs cheese in the United Kingdom. Both climates have relatively cool atmospheres almost year round. The lettuce for the big is grown in Spain due to the fact that it needs a warm climate to grow in. The lead item on the big mac is the onions and these are found in Holland. Holland has cool summe rs and mild winters which make it the optimal place to grow onions.Onions need temperatures to be at a happy medium in the midst of hot and cold for them to grow well. All of these places and items grown are promptly related to the McDonalds in the United Kingdom. The United Kingdom gets all of its ingredients for its big mac from different countries. The reason they do this can be described by Webers model. Webers model centers on three main ideas much(prenominal) as transportation, labor, and agglomeration. Transportation in todays modern world is fast and gaudy meaning that it is a viable option to ship products to different locations that may be far away.The fact that there is cheap transportation makes it easier for countries, such as the U. K. , to get products that they do not have the potential of growing as well. Labor is a much more important component of Webers model today. This part of the model is classified as the most costing part of an organization. This means t hat companies will look for a cheaper labor pool. The U. K. follows this pattern for the most part. The United Kingdom has a minimum rent of $9. 83 in U. S. dollars.Some of the main suppliers for the items in the big mac for the U. K.are Germany, America, Canada, Turkey, and Spain. All of these places have less of a minimum wage requirement than the United Kingdom. This means that it is cheaper for the United Kingdom to pay workers in other countries to grow their products rather than grow them in their own country.Lastly, there is the matter of agglomeration. Agglomeration is often when businesses that produce the same product locate around each other to reap some sort of benefit. This directly relates to all 1,200 McDonalds in the United Kingdom. Many of these restaurants are located around other fast feed chains.The main idea behind this is to potentially steal costumers from other fast food restaurants. McDonalds can also benefit from other fast food chains advertisements if t hey are located near them. McDonalds tends to symbolize development in countries. Every developed country in the world practically has a McDonalds. Most volume credit McDonalds for being the innovator of the development of the fast food industry. The food chain has developed a well-oiled machine for creating the same products. They have people located at certain areas to do specific jobs such as grilling burgers, adding dressing, frying fries, getting drinks, and cashiering.This may sound similar to another model that gave descent to the auto industry. This model being the hybridization model. Ford was the first to develop an assembly line, which basically gave workers a specific job that they only did. This maximized production for Ford made it so they could produce cars in a more efficient time frame lowering prices. Due to the fact that McDonalds runs their business similar to Fords is the reason why their prices are so affordable. Many different companies have shaped their bu sinesses around McDonalds. This is why there are so many different typesetters cases of fast food chains.Since there are so many McDonalds worldwide there are bound to be negative side effect centering on the issue of consumption. McDonalds food is very high in sodium and fats. This type of food is not very well-preserved for the average persons diet. This means that the food consumers are consuming at McDonalds can contribute to obesity if they eat there too frequently. The obesity rate in the United States is 35. 7% of all adults. These statistics show that obesity is a huge issue. There is shortly 550 million big macs sold in the United States each year. This is not a very healthy product.The fact that so many people are buying and eating it could contribute to their unhealthy weight. There is also a matter of how much virtual water is used to make the beef bar in a big mac. There is 4-18 thousand gallons of water used per beef patty in a big mac. The reason this number is so high is due to the fact that it takes a grapple of water to grow grain for cows to eat and cows drink a lot of water. body of water seems comparable a substance that can be found on a whim. However, it is actually a relatively hard thing to find. For instance, people around Texas are actually mining water.They are mining in the Ogallala Aquifer which is basically a huge underground supply of water. The fact that people have to tap water just adds to the fact that there is not an abundant amount to be wasted like is being seen on the production of a meat patty. There is also the matter with the idea that the more McDonalds that are spawned in certain areas there is less of a culture in areas. This is the idea of placelessness. Areas fall behind their own culture due to the fact that everything starts to look the same. The more demand for McDonalds means more stores and less of a cultural background in places.

Liberalism remains hugely influential in British Politics Essay Example for Free

Liberalism remains hugely powerful in British Politics EssayLiberalism remains hugely influential in British Politics, moulding the care for systems and policies of all the mainstream governmental parties. Discuss.The prime focus of Liberalism is freedom and liberty the type of liberty they strive for depends, however, on which of the two types of Liberalism is being referred to. Firstly classical Liberalism, believing in negative freedom and progressive Liberalism disputation more towards corroboratory freedom. I would argue that Liberalism does still have an influence in British political sympathies however I would non go as far as to say that it is hugely influential. Firstly, I would outline that the work companionship is possibly the least liberal in their political show up. Their tello does non set out many liberal policies due to their economical stance. The fact that Labour is traditionally a moderately socialist party, conpennyrating on equality indoors soc iety and for this reason, they would steer away from the free-market and limited government intervention approaches that liberals may take to politics.The limited government that Classical liberals see as ideal would not appeal to the Labour party as they strive for the equality of their citizens and would feel that they need to impose policies, such(prenominal)(prenominal) as their policy on housing set out in their 2010 manifesto No depression duty for first-time buyers on all house purchases below 250,000 for two years, paid for by a volt per cent rate on homes worth more than 1 million. This policy shows a socialist position to New Labour, highlighting a from each according to his ability, to each according to his need ideology within Labour politics.However, progressive liberalism may be seen to have had more of an influence on Labour Party politics. The fact that the Labour party believes in imposing policies of equality on society implies that they value overconfident f reedom. Policies put forward such as the Equality Bill which has been justified as a integrating measure bringing different strands of anti-discrimination law together while implementing EU directives highlight the controlled aspect of the Labour partys equality regime.It could be argued that in the past, aspects of Liberalism, although neither truly Classical or Progressive, such as Neo-Liberalism have had a significant influence on British politics as easily as the policies and value systems of the nonprogressive party in particular. During Thatchers time in office, she highlighted an ideal of a pragmatic means to progress. The Thatcherite Conservative party saw Neo-Liberalism as an obvious way to adapt to the endlessly changing character of modern political thought. Tony Blairs New Labour could also be seen as a Neo-Liberal third way government, on both a social and economic scale.The current Conservative party however, seem to be significantly less affected by Liberalism. Ar guably, the free-market ideals that Cameron appears to hold are seen as Neo-Liberal however, it is discernible that in the current coalition government that Britain is currently led by, this free-market and Neo-Liberal philosophy has been limited to the catch-all demands of mainstream politics.This is apparent in Conservative economic policies such as that put forward in their 2010 pre- planetary election manifest A five per cent pay cut for Ministers followed by a five-year freeze, and a 10 per cent reduction in the number of MPs. This policy in itself shows how Camerons Conservative party are stressful to appeal to the masses in controlling the pay of ministers. This does not inevitably denote a free-market approach to the economy and does not play into the meritocratic values that have been seen in the Conservative party in the past.It could also be argued however, that more prominent economic policies put forward in the Conservative manifesto pre-2010 have shown that Camerons Conservatives are moving toward the Neo-Liberal, Thatcherite ideology that the Conservative party once had. Although this may not have been the case when Cameron first became the leader of the Conservative party in 2005, aspects of his partys general election manifesto in 2010, such as Reduce welfare dependency and Stopping revenue credits to families with incomes over 50,000 have proven to have shown the partys shift back towards Thatcherite economic politics. These policies show a move back towards meritocratic society and a distinct step away from the something for nothing culture that has emerged in modern Britain.Finally, the Liberal Democrats both by name and by nature are arguably more influenced by Liberalism than any other mainstream political party. Progressive Liberalism can be seen clearly in certain aspects of their recent policies, on education for example Cutting class sizes to get wind children get the individual attention they need. This is a fine example of how t he Liberal Democrats would like to impose positive liberty and freedom on its citizens cutting class sizes would increase individual attention and therefore arguably create more freedom and liberty for pupils in the long-term.It could also be argued however, that the Liberal Democrats are no longer such a liberal party, but more of a third way political party much like the Conservative and Labour parties who have shifted towards the centre of political ideals and values in order to also become more catch-all and appeal to the masses.In conclusion, I would argue that broadly Liberalism is not hugely influential in British politics as a whole and does not necessarily mould the value systems and policies of all the mainstream political parties. However, I would argue that liberalism still remains influential in certain specific parts of each partys policies and value systems therefore remaining an influential force, to an extent, in British politics.

Contractors in the Military Essay Example for Free

Contractors in the Military EssayGoing game in time to the Civil War, private contractors can be found, providing retain, logistics, and supplies to the army forces. If you think about at presents military bases and approaching the front gate that do you fascinate? substantially the first mortal you see is a private security department guard standing in that respect waiting to welcome you to foregather whatever. Why dont you see a Military Police Officer standing there? Simple question right, well it isnt because the Military is deviation more and more towards a noncombatant work force. What does going to basically civilian work force do for those men and women that serve in the United States Military today? Is the judicature really making the right choice by go on to use private contractors back home and abroad in war z champions today? That is where this paper is going to search out the causes and effects of the use of private contractors or civilian work force i n todays Military. Todays Military bases are flooded with civilian workforce works for the government as private contractors.The Government today is looking for anyway to cut Military budget and save capital. But is using the civilian workforce saving money? Well lets talk about those security guards at the front gate, lets say the guard is amid a GS 1-3 the average recompense for them are seventeen thousand to twenty eight thousand a year. Now look at an Army Military Police Officer e-1 through e-3 which is equivalent to the GS 1-3 the salary for that e-1 through e-3 is between seventeen thousand to twenty thousand dollars a year. So from that little equation using a private workforce really isnt saving anyone any money. Let me inspire you that doesnt include years in service. Because more time in the GS ranks you make more money faster. But for that enlisted soldier his money caps after so many years. But everywhere you go you see more and more civilian workforce so there mus t be a corroborative font to having civilians work alongside with the Military agencies.Having private contractors work alongside with the Military can seclude some stress off of units that are deployed down range in combat zones. Those units can localise on their missions at hand and can rely on logistical support and personnel support from basically private security forces downrange. For the State Department, armed contractors likewise perform a variety of security tasks(Isenberg 2009). In comparison to past deviations the United States was involved in like Gulf War one in which the dimension was one contractor to fifty eight service men. Now lets jump to the Bosnia conflict where the ration jumped to one to fifth teen. Then you look at Iraq War which the ratio dropped to one to six. So the use of the contractor is becoming more evident in armed conflict zones. So many jobs that the Military gives to the private sector are specific jobs or a valuable skill that is a specifi c task. So by giving those specific jobs to contractors in war zones allows the military to provide sustainment abilities on forward bases.Then you spend a penny to look at the bad side of those private contractors, with the likes of Black Water, and Triple Canopy(Cotton 2010). Those two companies have been involved in many altercations that define a bad taste in peoples mouth about private companies functional in war zones. Each company did many things that caused a lot of crisis for fellow service members charm they were deployed in hostile places. These companies go to do a job get p back up more by our United States Government then goes and does hostile acts against the other country, that puts many other people not save service members in harm way. So why should the DOD pay people like this? Well that is just the great debate.So with the government in war still in Afghanistan, why do we continue to rely on so many private companies, well that is because of those vital asse ts those people offer. So if the government would put a little more faith in the Military Services we have and train. We could start working on the costs of what is spent while we are at war. Even though it would be great to limit the use of contractors, the use of the contractors just needs to be limited in certain ways. Keep using the ones that are needed to assist in logistical support and aid in missions to assist. Try and stop the use of the ones in war zones running missions could improve on the government spending, let those people that serve in the Military keep running those missions, because that is what enlisted do serve our country. industrial plant CitedISENBERG, DAVID. Shadow Force. Westport, CT Praeger Publishers, 2009. Praeger Security International Online. 1 Jan 2013.

The existence of high commitment management in unionized workplaces Essay Example for Free

The existence of senior luxuriously school loyalty concern in substanceized workplaces EssayThe existence of high riptedness centering in nonpolar workplaces suggests that much(prenominal) focusing is congenial with trade unionism. DiscussThe relationship betwixt workers and anxiety has historic tout ensembley been an adversarial one. In the process of stiring for employee right and benefits, unions endure traditionally created a them and us atmosphere surrounded by employees and managers. On the other hand, high committal way tries to eliminate the them and us atmosphere and instead attempts to switch over it with one of partnership where both employees and attention work together for the benefit of the firm. Thus, the respective natures of high dedication establishments and unions would imply incompatibility. It is due to this belief that some high commitment firms devote enormous resources to strategies aimed to keep unions outside from their firms such a s hiring lawyers who specialize in union avoidance and implementing high commitment work practices such as grievance systems and work teams.However, there is a paradox to this belief as studies show that as compared with non-unionized firms, unionized firms in high commitment establishments provide more than rights and benefits to workers, give more opportunities to employees to give and put one across information, and give a greater voice to employees to voice either constructive suggestion or differ (WERS 1992). Furthermore, union presence is increasingly popular in HRM organizations, as union density is 47% in organizations which employ at least half of the identified high commitment management practices. In this paper, I will use evidence from various studies to show why high commitment firms and unions are and then compatible.High commitment management is based on the theory that firms who give moreHigh commitment management is based on the theory that firms who give more to their workforce get back more from it in return. This requires a cooperative relationship based on trust and reciprocity mingled with intentness and management. Unions cigarette help in solidifying a high commitment environment by policing management to ensure that certain high commitment goals are met. Unions in high commitment work environments fight to give employees a collective voice in decisiveness making as well as enhancing suffusive and procedural justice while in return improving workforce solidarity and aiding in legitimizing pitch. Thus, the compatibility amongst management and unions can increase the payoffs of both management and labor while allowing the firm to firm to be driven forward.In the fast pace, ever changing corporate world, responsiveness to change and flexibility within firms are oftentimes required so that firm can readily adapt to ever-changing times. upstart times have seen an increasing amount of firms moving away from traditional authoritar ian ways of management towards that of high commitment human resource management. Such drastic changes in work practicess however, are non always easily implemented because a lack of communication among the workforce and managers makes it hard to change the character of employment relationships. Unions however, can aid in making such changes by allowing firms to communicate to the constitutional workforce quickly and efficiently while giving the workforce the opportunity to respond with insert or inquiries.This increases the workforces k straightledge and apprehensiveness of the new direction that their firm is taking and improves the probability that they will facilitate sort of than resist future changes. Such was the case with Tesco, who realized that its inflexible employee involvement structure, as well as its high turnover, relatively low pay and a hands on directive management dash were inconsistent with achieving enhanced levels of service to advance in competitive st rategy.1 Tesco lacked their employees to recognise with and commit themselves to the company, and it saw the union Usdaw as the means for cultivating and realizing this goal. As a result a partnership between Tesco and Usdaw was formed where Usdaw gave up their strike threat in return for a greater union input to secure improved policy implementations. As a result, employee wages rose, terms and working conditions were improved and ply turnover dropped significantly.2 . Thus, this partnership took a step away from an us and them environment and took one towards unions working with management as team for the benefit of the entire firm.High commitment management is greatly dependent on a partnership between employees and management. This assumes that there arent agency problems because it is expected that employees have a deep understanding of the firms interests and that they go about their job as if those interests were their own. In order for the partnership between employees an d management to flap however, a workforce must have a collective voice in decision-making. Firms encounter many problems when trying to give individual workers decision-making authority. For instance, there is always the fear that a worker will revilement his decision-making authority by using it to further his own ends rather than to help the entire firm advance. other problem is that it is extremely difficult to have a partnership between management and a abrupt workforce. Unions however facilitate a partnership by providing a channel of communication between management and the entire workforce.3 This enhanced communication is made possible by unions because just as management speaks with single voice, having a union allows the employees it represents to speak to management with a unified voice. Thus unions give their members decision making authority by taking into account the needs of the workforce and communicating it to managers. As a result, there is an improved match bet ween what employees want and what they get from management and this is an important ingredient in high commitment environments.Allowing workers to have a say in how their firm is managed increases their job satisfaction and is a major reason why on average, turnover is lower in union settings.4 In addition, unions bleed to fight more for the benefits of employees with longer tenures in firms. This gives union members an increased incentive to commit themselves to their firm in the long term and also may be a factor contributing to lower turnover. This is compatible with high commitment management strategy in many occupations such as teaching and nurse where goals such as low turnover and worker longevity are important.However, any high commitment based partnership can break down if there is a lack of trust and accountability. Unions though can be trusted by management to act cooperatively because managers know that union officials want to protect their unions reputation with a par ticular firm and oftentimes this concern for its reputation can be the guarantee that management needs of good, constructive sort5. In addition, a long-term partnership between unions and management can permit a greater degree of trust and objectivity than can the partnership between management and individual workers. This is because there is a greater frequency of interactions between management and a union.The partnerships find out in collectively dealing with past disputes makes it possible for future disputes to be settled in a promptly in a way that doesnt disturb work practices. This partnership also presents a checks and symmetry system to ensure that both parties act cooperatively. Just as a union is a caller that can be held accountable for the actions of its members6, management can also be held responsible for bad behavior of any single manager. For instance, if a manager is engaging in an activity that the workforce views unfavorably, a union representative can give notice to management to put an end it before it goes too far. matchless of the biggest obstacles not permitting employees from fully committing themselves to a high commitment firm is if they perceive there to be inordinate unequal treatment. This can hang employee performance incentives as well as hurt the morale of a high commitment firm. Unions however, have a sword of justice effect and are successful in eliminating many of the adverse affects that occur because of unequal employment opportunities. A major reason for this is that unionized can better monitor firm practices to ensure fairness because they are much more credibly to have managers and supervisors who are trained in people management skills and in addition, managers in unionized firms put a greater emphasis on ensuring that there is equal treatment among their employees.Studies indicate that when taking into account workplaces where all the supervisors are required to have training in people management skills, 20 % of those in unionized environments, as compared with only 12% of those in non-unionized places, have responsibility for equal opportunities.7 Two of the most common and document forms of unequal opportunities are employee promotions and employee selection. When compared to non-unionized firms, firms with a union presence are four times as likely to monitor promotions by gender and three times as likely to review selection procedures to identify indirect discrimination.8. There is also evidence that the increased monitoring in unionized firms has helped reduce inner discrimination as it regards to promotions and this has thus helped women advance up firms hierarchy.Studies indicate that about one in seven-spot managers in the union celestial sphere reported that the proportion of women in the managerial post had risen markedly in the last 5 years while only one in ten managers in the non-union sector reported this. 9 Furthermore, unions have also traditionally been seen as defe nders of an egalitarian pay structure. High commitment work environments value and oftentimes even require worker solidarity. However, when some workers are paid more than others for essentially the same job, the existence of this solidarity is difficult to maintain. A union presence in a high commitment work environments facilitates the presence of solidarity as they prefer to pay each occupational group a single rate.This takes the wages out of competition among co-workers, and ensures that the high-commitment working environment wont be harmed due to unkind competition practices such as undercutting between co-workers. Thus unionized workers have less wage difference than their non-unionized counterparts. Studies indicate that the standard deviation is .457 in unionized setting while it is .592 in a non-union setting.10 Furthermore, unions help decrease wage inequality by 2.6% between males and females, 1.4% between blacks and whites, .6% between the healthy and those with healt h problems, and 3.1% between non-manual of arms and manual employees.11. Thus by ensuring that firms pay equal wages for equal work, unions help maintain workforce solidarity which acquired immune deficiency syndrome firms in maintaining high commitment work environments.In recent years, it had become apparent that the relationship between firms and unions doesnt necessarily have to be an adversarial one. In fact, the popularity of partnerships in high commitment firms indicates that the two institutions are and so compatible. The evidence stated above indicates that when this partnership is properly utilized, unions can be a great plus to a high commitment management work design. This is because unions facilitate the existence of many of the necessary ingredients that go into high commitment work environments such as flexibility, increased employee voice in decision making, distributive justice, and worker solidarity. Major firms such as Tesco have recently taken advantage of thi s compatibility and are now realizing the formerly untapped benefits that a partnership between unions and high commitment management schemes can shape about.1 Industrial Relations Services Partnership delivers the goods at Tesco, IRS Employment Review, August 1999, No. 6622 Industrial Relations Services Partnership delivers the goods at Tesco, IRS Employment Review, August 1999, No. 6623 Fernie, Sue. HRM yack away on 29/01/02 London London School of Economics, 20014 Baron, James N. , Kreps David M. strategical Human Resources Frameworks for general Managers. (New York John Wiley Sons, Inc., 1999), 1275 Baron, James N. , Kreps David M. Strategic Human Resources Frameworks for General Managers. (New York John Wiley Sons, Inc., 1999), 1346 Baron, James N. , Kreps David M. Strategic Human Resources Frameworks for General Managers. (New York John Wiley Sons, Inc., 1999), 1347 Fernie, Sue Gray, Helen Women what unions can do for you centrepiece Summer 20008 Fernie, Sue Gray, H elen Women what unions can do for you centerpiece Summer 20009 Fernie, Sue Gray, Helen Women what unions can do for you CentrePiece Summer 200010 Metcalf, David Fighting for equality CentrePiece Summer 200011 Metcalf, David Fighting for equality CentrePiece Summer 2000

Role Of Perception Essay Example for Free

Role Of Perception EssayPerception plays a vital role in a beings life, especially in that of a human. Many people choose to opine what they see, and accept cogitate with knowing as it is a component of knowing. We also choose to see what we trust and therefore both statements argon complimentary.Human beings tend to believe with evidence rather than believing in something blindly. Belief is ordinarily based on evidence and the evidence is usually linked with our senses. If we see something, we believe in it because we be able to impale our assumption with evidence because we have witnessed it. Human reality whitethorn also be based on scientific evidence because science which involves historical evidence is able to prove au pasttic theories. redden though we may not witness it with our senses, we believe in the theories accepted by umpteen other scientists. For example, the world trade center was demolished on September 11th, 2001. We suddenly think, how is it possible to demean one of the greatest and tallest buildings in the world even after the first attempt to destroy it back in 1993. Suddenly as we see it constantlyywhere in the news and read it everywhere, we strongly believe in it rather than half believing in it because we have seen images and video clips of the incident.Humans, according to E.F. Schumacher have the elements of matter, life, consciousness, and self aw areness or intelligence. Our self awareness modifys us to see things the way we do and it is accepted psychologically that we hold fast to what we can see and experience.Yet knowledge plays a vital role in believing along with row. Since we can read and understand as well as communicate, we can percent ideas expanding our knowledge. This also ties in with the senses of hearing and seeing. Knowledge according to Plato is justified true(p) vox populi. Our senses of hearing and sight enable us to justify what we believe. Since elementary we have learned simple to comple x equations and yet we are chill out learning. Some smart mathematician discovered that 2 + 2 equals 5. We have seen for ourselves by taking two pieces of drinking chocolate for example and adding another two pieces of chocolate we get tetrad pieces. A simple experiment much(prenominal) as that enables us to see as well as learn simple mathematical equations. This authentically helps create a basic foundation in learning, especially in the mind of a child. that if it were not for language, 2 + 2 superpower as well equal a parasite or two thousand if it were not for language. We have learned the basics of English or whatever language we know and therefore it makes sense to us that 2 + 2 equals 4 rather than any fare which we exponent consider ridiculous.It is also very easy to be fooled into seeing what we believe. It is both good an drab in a sense only it depends on the person. I am referring to people because we have four elements matter, life, consciousness, and intelli gence or self awareness- according to E.F. Schumacher. Animals lack self awareness according to Schumacher even though they have some intelligence. I think it is true because have if we think about it, have we ever seen a dog making a computer or bombs? In the most extreme human face a dog may be able to move a mouse or open air a program or maybe short a bomb and make it explode, plainly then again others might object because these thoughts may vary from person to person according to their point of view.We are sometimes forced to believe certain things without having 100% evidence showing or stating that its true. Take piety for example. I believe there is a god but how do we choose when there are over millions in the world. I believe that there is a god controlling the universe but how do we know for sure? I believe that is where faith comes in. Sometimes we have to believe blindly.Even though in the Bible it says In the beginning god created heaven and earth and saw that it wa s good, it is considered a mythical story. Who saw god create earth, light, water, or animals? According to the Bible, Adam was created on the 6th day, so there is no way a human took an account of the events as they happened. In religion, we are natural into a tradition which enables us to see through rituals. It is a framework to many which leads them through the joys and troubles of life. piety can be a new way of seeing, however it may create havoc by causing people to jump to conclusions. For example, a person was involved in a car accident. think they were traveling at a high speed and they lost controlledand crashed into an oncoming truck. The car is totaled but the person is knocked unconscious but returns to consciousness realizing what has happened and they have to broken bones or scratches indicating what has happened. People may immediately say it was by the grace of god nothing happened to me, but if we think about it, it is their belief that they are seeing no matter how hard you try to convince them it was something else.A simpler example would be that of a person who is terrified of ghosts. This person might believe in ghosts robustly and then might think that they have seen one in the middle of the night. What most probably has happened is that this person believes strongly that ghosts endure and what happened was that they saw a fluttering white sheet or a moons shadow which has been cast on a scarecrow which is swaying in the wind. The mind immediately plays a trick on this person, fooling them into believing that they ad seen a ghost.Another example could be a man who cannot see beauty. He might not believe in it at all and finally when the most beautiful woman in the world crosses him, he may not be able to realize her beauty. These two examples can be examples of when seeing what you believe may fool a person.I think the statement you see what you believe is true because we do tend to believe entirely if we have some sort of proof or e vidence. You see what you believe is also true as we tend to believe in things blindly as well. I feel it may be misleading at times but the statement is true and this partial statement completes the first.

High School Dropouts Essay Example for Free

high school School Dropouts EssayIn The Great Gatsby property is a symbol of success. Money controls the lives of the characters in the story, tho it is easy to see that it could not bring happiness. Each of the characters thinks that having money will solve tout ensemble of their problems and derive them successful, but that is not true. These are a few examples of characters in the story who think that money is the break up to being successful, when it actu each(prenominal)y does not bring them true happiness. Daisy is a character who was born into a affluent family, but she does not have any good encourages or purpose in her conduct. Her life is boring, so she tries to use her wealth to make her happy. She is a socialite, eer goes to parties with her wealthy friends, and she wears clothes that are clean-living with a lot of gold and silver.Even though she thinks that these things will make her happy, she is still bored and always wonders what she will do next. She h as an affair with Gatsby even though she is married to a wealthy man, Tom, because she is bored. She does not value herself or anyone else. Her money has not made her a happy person even though everything she does is base on wealth.Gatsby has a lot of money. Even though he has a huge house, fancy cars and clothes, and has parties all the time, he is not veryly corrupted by the money. He does everything he does just to grab Daisy and be to her by his wealth that he is good enough for her. He thinks that all he has will get her to marry him, but he does not realize that Daisy will never leave her husband because he is so wealthy.Daisys husband Tom is very wealthy. He gambles, goes to horse and car races, and has many affairs. He has a mistress that he supports, but obviously not happy with his life. When he finds out that Daisy has been having an affair, he gets angry with her even though he is guilty of doingthe same thing. Daisy and Tom are not happy, their money and success hav e not made them happy, but daisy will not leave Tom because he is wealthy.In The Great Gatsby money is seen as a symbol of success by the characters. They have all of the clothes, cars, houses, and social events that they want and that money can buy. Despite all of this, they are unhappy, have no real values or friends, and no real purpose in life.

Isolation of Recombinant Escherichia Essay Example for Free

isolation of Recombinant Escherichia Essay unmatched technique important in both elementtics and biochemistry is the Polymerase chemical chain Reaction (PCR), first developed in the 1960s, and thusly automated in 1983. Current PCR technology was non developed until the discovery of thermostable polymerases, particular proposition tout ensembley Thermus aquaticus (Taq) polymerase (1). The protein Taq polymerase was first discriminate from the extreme thermophile T. aqauticus, where extreme thermopiles argon bacteria that locomote in temperatures at or above 45C. The Taq enzyme is a member of the DNA polymerase I family (2, 3). The elicit property of Taq polymerase is that it has a temperature optimum at 74-75C, tolerateing it the remain active in temperatures required for PCR double quarantined DNA denaturation (3, 1) . The protein has an approximate molecular weight of 6263 kDa when insulate from T. aquaticus, and 94 kDa when isolated from recombinant Escherichia col i, and is still stable at temperatures of 93-95C, hence the thermostability of the enzyme ). Taq specifically deficiencys any proofreading activity in the 3 to 5 direction, and accordingly has a comparatively high error rate of single brutish mispairings of 1 error perIsolation of Recombinant Taq Polymerase for PCR 9000 nucleotides, as fountainhead as a frame shift error rate of 1 per 41,000 basepairs (5, 6). Taq polymerase has an activity that is highly st contrisolelyeg- push with on the environment of which it is in as it is thermostable, and has differing activities at nearly all temperatures up to the point of denaturation. Taq specifically digest add up to carbon0 base pairs in length on a scout in under virtuoso minute under typical PCR conditions. The enzyme has a specific activity of 200,000 units mg-1, and faeces add approximately form 60 nucleotides per second at 70C (7). The isolation of Taq is demand for the PCR reaction. The most important reason for Taq bei ng dropd in PCR is the thermostability at high temperatures (95C). This willed for the process of elongation, annealing, and denaturation to kick the bucket without the replacement of saucy enzyme, and thereby, was more efficient, faster, and cheaper beca subroutine the reaction could be automated through with(predicate) the use of a utensil known as a thermocycler which basically is just a machine able to change temperatures of an isolated environment rapidly (7).Prior to the discovery of Taq, PCR was do victimization Klenow fragments of E. coli DNA polymerase I at 37C. The lack of thermostability required replenishment of enzyme after(prenominal) apiece PCR cycle (8). wholeness of the initial difficulties of Taq polymerase was the organism in which it was expressed in, T. aquaticus, as it was difficult to civilisation and produce large quantities of enzyme. E. coli bacteria were engineered to expressed the Taq polymerase gene to allow for retrieval of large quantities o f enzyme ). The isolation of the Taq gene involved culturing T. aquaticus and accordingly isolated the DNA of the cells through lysing, protease K addition, extracting of aqueous and phenolic phases, dialyzing of extractions, addition of SDS, and then centrifugation of closure to eventually retreieve the DNA of the organism as delineated in Lawyer et al., 1989. With the isolation of the 2401+ BP gene of Taq, the gene was incorporated into a 6.58 kbp plasmid (pLSG1). The gene was inserted 171 bp distal to the lacZ promoter/operator, and 109 bp distal to the BgII site, so the gene expression could be controlled through an inducible promoter.With the pLSG1 plasmid, the vector was introduced to E. coli bacteria to allow for plasmid uptake (4). Other try outs throw away been conducted towards the subtlety of Taq from recombinant E. coli. Specifically En mousseke et al., 1990 developed a order for purfication of Taq. The E.coli strain 2 DH1 was used for the expression of the recom binant plasmid containing Taq polymerase. The bacteria were gravid in 12 cubic decimetre batches of Luria Broth using 1 mL of saturated DH1 culture and 80g/mL of ampicillin. Isopropyl-1-thio--Dgalactopyranoside (IPTG) was added to 0.5mM and the cultures were grown for 16-20 hours. The cells were harvested in 2.4 L of cushion A (50 mM TrisHCL, pH 7.9, 50 mM dextrose, 1mM EDTA) and collected via centrifugation, re freeze downed in Buffer A with 4mg/mL lysozyme and incubated at manner temperature for 15 minutes. Buffer B (10 mM TrisHCl, pH 7.9, 50mM KCl, 1mM EDTA, 1mM phenylmethylsulfonyl fluoride (PMSF), 0.5% Tween 20, 0.5% NP-40) was added and incubated in 180 mL fractions, for 60 minutes at 75C in a water bath.The mixtures were centrifuged at 8000 rpm for 15 minutes at 4C. Taq then precipitated with polyethyleneimine (PEI) at mode temperature, then isolated through centrifugation and suspended in buffer C (20mM HEPES, pH 7.9, 1 mM EDTA, 0.5mM PMSF, 0.5% Tween 20, 0.5% NP-40) c ontaining 0.25 M KCL. PEI eluatents were diluted in 50mM KCL and buffer C and applied to a 150mL BioRex 70 ion exchanger column, and then eluated using 200mM KCL. The protein was dialyzed for 12 hours against two changes of 1 L storage buffer (20mM HEPES, pH 7.9, c mM KCL, 0.1 mM EDTA, 0.5 mM PMSF, 1mM dithiothreitol, 50% glycerol.The experimentation resulted in 40-50 mg of protein per litre of cell culture (9). The methods used in this experiment differed in certain key aspects. First, Engelkes experiment make use of a high concentration of ampicillin. The IPTG was added to the like concentration, but was added after cell growth up to an visual density of 0.700. Instead of a water bath at 75C, this experiment do use of an air incubator for the temperature requirements. Engelkes experiment made use of PEI to precipitate Taq, while this experiment made use of 30g of (NH4)2SO4 per cmL of supernatant. Buffer C was not used throughout this experiment, and no ion exchange columns were used. The dialysis procedure was d angiotensin converting enzyme for twice as long with twice as many changes of ascendent per 6 hours. The changes made from Engelkes experiment offers a different method for protein ruin. The method used by Engelke made use of PEI which is an relation precipitation method versus a salt precipitation method.The PEIIsolation of Recombinant Taq Polymerase for PCR method has the major drawback through the lack of selectivity, and can often precipitate nucleic vinegarishs as well (10). This is why the BioRex column needed to be used. ammonium sulfate has the advantage that the precipitation can be controlled based on ionic strength of species involved, as well as has no negative effects on the activity of the target enzyme. Salting out also has the advantage that only native tell apart proteins are precipitated collectable to the hydrophobicity involved with native state proteins (10). Buffer C was not required for this experiment as no BioRe x column was required. This experiment made use of various techniques and methods including SDS-PAGE, differential centrifugation, horse opera Blotting, real time-PCR (rtPCR), PCR, agarose gel electrophoresis, and dialysis. dickens important techniques were PCR and rt-PCR. PCR does not allow for the quantification of DNA amplicons as it is an end-point PCR, but it does allow for confirmation of template duplication along with measurement of base pair length. Amplification of primer would confirm the presence of a thermostable DNA polymerase.The following agarose electrophoresis suspensors to amaze amplicon size which can tell us the activity of Taq, as well as the specificity, as sensation(a) template should only return one lap in PCR (7). rt-PCR allows for a quantitative assessment of PCR, and then the kinetics of the reaction, as it detects the add of amplicons produced in the reaction. The point at which the stock nose reaches threshold in cycle number gives informati on on the activity of Taq, as a more active sampling of Taq reaches threshold earlier.Melt curve synopsis also provides information regards DNA amplicons in resolve (11). The conclusion of this experiment was the test the methods for the isolation of PCR grade Taq polymerase from recombinant E. coli using differential centrifugation, salting out, and heat denaturation following lysation of cells to potentially improve isolation of Taq from past methods. The presence of Taq will be confirmed through western blotting, and rt-PCR and PCR reactions along with award will be assessed through SDS-PAGE. The activity of Taq will be found through rt-PCR and PCR. decision the most efficient method for the isolation of Taq offers a valuable reagent source for any PCR reactions required. The isolation technique would also be applicable to any thermostable proteins. 3EXPERIMENTAL PROECDURES Isolation of Taq Polymerase Luria broth (500 mL + 100g/mL ampicillin) was inoculated with 50 L of fro zen Taq polymerase expressing E. coli cell stock. Incubation was commenced for 12 hours at 37C until the Optical Density had reached 0.700. IPTG (0.5 mM or 0.112g/L culture) was added and the culture was incubated for 12 to 14 hours at 37C. The 50mL of cells were then centrifuged (4000 RPM x 15 minutes at room temperature) in an Eppendorf centrifugate 5810, and 5 mL of buffer A (50 mM Tris-HCl, pH 7.9, 50 mM dextrose, 1mM EDTA) was used to suspend the separated pellet. The solution was then centrifuged again (4000 RPM x 15 minutes at room temperature) in an Eppendorf Centrifuge 5810 and the pellet was once again suspended in Buffer A, with an special 20 mg of lysozyme added. The reaction was incubated for 15 minutes at room temperature.Following incubation, 5mL of buffer B (10 mM Tris HCl, pH 7.9, 50mM KCl, 0.5% Tween 20, 0.5% NP-40, 1mM PMSF, 1mM EDTA) was added and incubated at 75C for 1 hour in a unseasoned Brunswick Scientific-Innova 40 incubator shaker serial, and shaken b y hand approximately every 5 minutes. The solution was then centrifuged (15000 RPM x 10 minutes at 4C) in a Thermoscientific Sorvall RC 6+ centrifuge and using a 603s Delta Range 30g of (NH4)2SO4 per 100mL of supernatant (8 mL of supernatant equivalent to 2.4g (NH4)2SO4 ) was added and incubated for 10 minutes at room temperature and shaken on the Innova 40 incubator.The lysate was then centrifuged again (15000 RPM x 10 minutes at 4C) in Thermoscientific Sorvall RC 6+ centrifuge and the resultant pellet was suspended in 2mL of buffer A. The solution was then dialyzed in a Spectra/Por tissue layer tubing set at 6000-8000 Da molecular weight selection in 1 L of storage Buffer (50 mM Tris HCl, pH 7.9, 50mM KCl, 0.1mM EDTA, 1mM DTT, 0.5 mM PMSF, 50% glycerol) for 24 hours at 4C changing the buffer every iii hours. The dialysis solution was then diluted in a 11 ratio of storage buffer and stored at -70C until needed. Protein Concentration Determination A Bovine Serum Albumin Bio-Rad ass ay measuring curve was nimble (0 0.3 mg/mL) usingIsolation of Recombinant Taq Polymerase for PCR a 1mg/mL stock solution and an Asys proficient Plus spectrophotometer set at 620 nm. Bio-Rad assay was run in triplicate using 20L of protein dilution and 150 L of diluted Bio-Rad Dye Concentrate. 10x and 100x dilutions of the savor prepared previously were made and 20L were used with 150L of diluted Bio-Rad Dye concentrate. The solutions were incubated for 10 minutes and absorbances were tabulated. sandwich was then assembled with an additional ice block in the transfer apparatus. The apparatus was run at 180mA overnight in a refrigerator and the membrane was then stored in TBST buffer (20 mM Tris-HCl, pH 7.6, 137 mM NaCl, 0.1% Tween 20) and refrigerated. 24 hours earlier to the primary antibody (Anti-Taq monoclonal Antibody (8C1)) addition, the membrane was blocked in 1 gram of Carnation nonfat ironic milk (5% w/v) and 20 mL of TBST Buffer.The primary antibody in TBST with SDS-P AGE 5% w/v nonfat dry milk at a 1800 dilution of A discontinuous polyacrylamide gel was antibody was added to the membrane and shaken prepared using a Mini-PROTEAN Tetra Cell for 1 hour at room temperature. The membrane was module. The stamp stand was assembled for 1mm then washed deuce-ace successive times for 15 minutes gel and fill up with National Diagnostics 12% with TBST buffer at room temperature. The Resolving gelatine (Protogel 2400 L, Resolving Gel secondary antibody (Peroxidase-conjugated Buffer pH 8.8 1560 L, dH2O 1974 L, 30% w/v AffiniPure Goat Anti-Mouse IgG (H+L)) was then APS 21L, TEMED 6L), casted to 1 cm below top applied in TBST with 5% w/v nonfat dry milk at a of glass plate, and then 4% Stacking Gel (Protogel 12000 dilution of antibody and shaken for one hour 390 L, Stacking Gel Buffer pH 6.8 720 L, dH2O at room temperature.The membrane was then 1830 L, 30% w/v APS 6L, TEMED 3L) was washed three successive times for 15 minutes with casted on top with a ten w ell gel comb. The TBST buffer at room temperature. The edges of the electrode set up was then submersed in 1 x Trismembrane were dried with a Kim and adjoining 1mL of Glycine SDS Page Running Buffer. To each 20 L Renaissance Western Blot kit out NEN Life Sciences of sample, 20 L of Laemmli buffer (0.5 M TrisProducts, Cat. No. NEL 101 luminol reagent with HCl, pH 6.8, 4.4% w/v SDS, 20% v/v glycerol, 2% 1mL of oxidizing reagent were mixed together and v/v 2-mercaptoethanol, 10 mg/mL bromophneol then applied to the membrane. The membrane was blue) was added and boiled for 3 minutes and then then imaged with an depiction time of 40 seconds cooled for 5 minutes on ice. To one well 7L of using AlphaEaseFC software. New England BioLabs Inc. Prestained Protein Marker (7-175 kDa) was added. To the following PCR/agarose gel electrophoresis wells 15 L of prototype Taq polymerase was added, A master mix for PCR was prepared (1x PCR along with 20L of six different samples, with the buffer e lectronegative Mg, 0.2mM dNTP, 1.5 mM MgCl2, fifth being prepared the previous year with the same 0.5M forward primer, 0.5m Reverse Primer, 0.1 method of isolation as outlined previously. The gel ng Template DNA and Nuclease-free PCR water) was run at 200 Volts for 40 minutes, incubated in and 22.5 L of master mix and 2.5L of Taq fixing solution overnight and then stained with Bio- sample, the standard, or the Taq prepared a previous Safe Coomassie Blue for one hour at room year were added to PCR tubes and centrifuged temperature under agitation.The gel was then briefly on a Fisher Scientific Accuspin micro 17 analyze used AlphaEaseFC software. just briefly using 1.5mL eppendorf tubes with no caps to contain the PCR tube. The PCR tube was Western Blotting then added to T3 Biometra Thermocycler and Using the method described above for SDS modifyd at 94C for 3 minutes and then 35 cycles PAGE, a SDS-PAGE gel was taken prior to fixing. of PCR with the denature 94C for 45 seconds, The ge l was then transferred to transfer buffer anneal 55C for 30 seconds, and extension at 72C (20mM Tris-HCL, pH 8.0, 150 mM Glycine, 20% for 1.5 minutes. The sample was then incubated at Methanol). Immobilon-P transfer membrane with 72C for 10 minutes and then temperature was 0.45 m pore size and Whatman paper were cut to maintained at 4C. The samples were then stored at the size of the gel. The membrane was wet with -20C until agarose gel preparation. A 1% agarose 100% methanol, then transferred to MilliQwater gel was prepared through 1.5g of agarose (Sigma and soaked for several minutes. A standard blotting No. A-6877 Type II) to 150mL of Tris-Acetate4Isolation of Recombinant Taq Polymerase for PCR EDTA (TAE) buffer. The solution was microwave oven for 1 minute and mixed until in solution. Once cooled to 60C, 7.5 L of Biotium Gel blushful Nucleic acid stain was added and mixed. The solution was then poured into the electrophoresis tray a comb was installed, and set at room temperatu re. One Litre of 1x Tae buffer was prepared through dilution of 50x TAE buffer and then the solution was poured onto the electrophoresis tray to cover the gel in 1mm of buffer. 20 L of PCR product prepared previously and 4L of Gel Red dye were mixed and 20L of each sample, the standard, and Taq prepared the previous year and Invitrogen life Technologies 1 Kb DNA ladder Cat. No. 15615016 was run at 150 Volts, 100 mA for one hour (or until dye reached the bottom of the gel). The hatfuls were then visualized under 300 nm light and fluorescence was metric at 590 nm.The gel was analyzed using AlphaEaseFC software. concentration of the sample Taq was 1.88 + 0.11 mg/mL. The solution of proteins was not fresh Taq as confirmed by the SDS-PAGE (Fig. 2) as various proteins created distinct bands (B to K excluding E). The standard Taq revealed only one band (A), indicating band E was most likely belong to Taq, as it was the darkest band in the gel. An analysis of the molecular weights of the bands through electrophoretic mobility (Tab. 3) showed the standard Taq having a molecular weight of 115.2 + 14.6 kDa, and the likely band (E) had a molecular weight of 113.4 + 14.3 kDa. on that point was a distinct distortion in the bands of the SDS page in all lanes with the exception of the standard Taq and the 2011 Taq (Fig. 3). The distortion is of a smile. The overall gel also has a large distortion, but of a frown. It would appear there was a similar protein to D E and F present in all samples, including the 2011 sample. The standard Taq did not contain the bands. Real Time PCR The Western Blot (Fig. 4) revealed distinct A master mix for PCR was prepared (1x PCR bands however, there were more than one band in buffer negatively charged Mg, 0.2mM dNTP, 1.5 mM MgCl2, each lane with the exception of the standard Taq. 0.5M forward primer, 0.5m Reverse Primer, 0.1 Two distinct bands were present in 5, Taq, and 2 (b, ng Template DNA and Nuclease-free PCR water). c).The lanes of * and ? contained several bands To PCR tubes, 22.5 L of nobleman Mix and 2.5 L also. The overall gel also expressed a slight color of Taq sample or the standard Taq were combined, stripes along the solvent front edge which is mixed through vortexing and then centrifuged with shown in both Fig. 3 and 4. The 2011 lane did not a Fisher Scientific Accuspin micro 17 just briefly appear to have any Taq present, as no band was using 1.5mL eppendorf tubes with no caps to distinguished. The entire ladder expressed some contain the PCR tube. The Taq samples were antibody activity. prepared in triplicates. 20L of each sampled were The real time-PCR revealed a threshold reached then transferred to a 96-well PCR plate and then at 20 cycles, with the colossal majority occurring at 24 sealed. The well was then placed in a BioRad CFX cycles. The melt curve showed an approximate connect Real Time System using the programing of melting temperature of 81C (Fig. 7). enzyme activation (95C, 30 seconds , 1 cycle), 40 The agarose gel electrophoresis revealed one cycles of Denaturation (95C, 1 second) and distinct band at approximately 5883.5 base pairs in annealing/extension (60C, 5 seconds), with a melt length.The brightest bands, and therefore the curve of (60-95C in 0.5C intervals, 3 seconds per highest quantities of Taq enzyme were found in the trample, 1 cycle). The samples were then analyzed std., 2 and 4. When the base pairs specific activity using AlphaEase FC software. of the enzyme was calculated it was found to be 834.5 + 63.9 bp/min/g of sample, or 3922.3 + 192.9 bp per minute. RESULTS The results of the Bio-Rad assay on the sample of Taq polymerases diluted to 10x and 100x revealed that the 10x dilution was far to concentrated and fell outside the analog curve of the Bio-Rad assay. The retrieval of protein from the Luria broth was found to be 300.8 + 17.7 mg protein per L of Luria broth. These results (Tab. 1) suggest the protein 5 tidings Through the analysis made through SDS-PAGE, the MW of the standard Taq was found to be 115.2 + 14.6 kDa and 113.4 + 14.3 kDa. This is different from the veritable literature value of 94 kDa (9). Even with error correction, the protein did not fallIsolation of Recombinant Taq Polymerase for PCR within the range of the accepted literature value. In total, the two proteins differ by 23% and 21% without error correction, or 21.2 kDa and 19.4 kDa noteively. In comparison to one an other(a), the two bands have essentially the same molecular weight, indicating whatever error occured in the gel was equivalent on both the standard and the isolated Taq. One explanation for the difference in the molecular weights may be explained through the quantitiy of protein used. The darkest and thickest band ( E, fig. 4) likely belongs to the Taq protein. To get a more defined band, a dilution would be effective in making a higher resolution band (12). The amount of protein isolated per volume of Luria broth was determined to be 300.8 + 17.7 mg per L of Luria Broth. Quite obviosuly, there are issues both with the heat of the gel, and distortion of the bands into smiles. The distoration of the gel likely was caused by unequal heating of the gel cause the center of the gel to be hotter than the peripheries, as the walls of the apparatus act as heat sinks (13).The uneven heating can be removed by switching to a lower voltage for a long-lasting period of time (12). The distortion of the protein bands within the individual lanes produced a smile structure. The distortion was likely caused by either an overloading of proteins, which can be solved by dilution of the protein sample, or was due to salt conditions of the loading sample. This pure tone could be fixed through extra steps of dialysis to decrease salt content of the loading sample. (14). One final issue with the SDS-PAGE gel was the distance between bands. The target molecular weight was near 100 kDa, so the concentration of the gel could be dec reased to allow for a higher resolution of the higher molecular weight proteins, or allowed to run for a longer period of time (14). A purity assessment of the isolated Taq enzyme can be made through the SDS-PAGE gel (fig. 2). Distinct banding occurs in ten different bands on the Taq lane, with 9 being distinct from Taq protein (E).This highlights that there were infact multiple proteins still present in the Taq solution. This would indicate that the heat shock portion of the methods was insufficient in denaturing all of the proteins in the E. coli, allowing for precipitation upon salting out. This is based on the extra protein banding only occuring for the Taq polymerases prepared for this experiment. A factor that could have also played a role was the incubation at 75C was continually 6 interrupted through the need to shake the reaction vessel thereby lowering the temperature of the solution. This was due to mechanical difficulties of the equipment. It would be best to find a wor king New Brunswick Scientific-Innova 40 incubator shaker series to improve the protein isolation. To decrease the protein impurities, an increased heat cycle could be implemented, as Taq is thermostable at 75C, and could lift structure at that temperature for long durations (7). The ammonium sulfate salting out would be more efficent after an increased heat cycle as even fewer native state proteins would remain (10). other method to decrease impurities would be to add a purification step using another specific property of Taq polymerase. This could be the isoelectric point.This could be done through ion exchange columnsor isoelectric focusing (12). The extra isolation step would significantly decrease the impurities, and increase the specific activity per mg of protein of sample.The impurities were likely a result of other proteins present in E. coli bacteria lysate that were carnal knowledgely thermostable, as those proteins would be most probable (9). The isolation of Taq can b e confirmed through the Western Blotting and PCR reactions (Fig. 4-7), as a distinct band in the Western Blot, and measureable amplicon replication in the PCR and rt-PCR. In the standard of Taq of the Western blot (Fig. 4) there is a distinct band. The same band in the channel containing the isolated Taq can be seen. The band occurs in the same relative vicinity as the Taq molecular weight band in the SDS-PAGE (Fig. 2) so would cash in ones chips best fit the Taq enzyme.The banding of the blot shows a common band across all lanes that line up with the standard Taq, emphasizing the isolation of Taq. There is a hesitation in confirmation of Taq due to the extra protein banding in the prepared fractions, as these bands were not seen in the standard Taq. The banding would suggest proteins transferred from the gel to the membrane and was still able to bind to the primary antibody or secondary antibody. There are various workable explanations for this. First and foremost, the banding oc curred in areas wherever protein was present (ladder and lanes). This would indicate lack of specificity in the primary antibody which is mean to only find full sequence Taq and bind to it (15, 16). Another problem may be due to lack of blocking solution binding to the membrane, or Isolation of Recombinant Taq Polymerase for PCR excessive laundry removing blocking solution from the membrane. A final possible explanation may be binding of the secondary antibody to membrane outpouring proteins with the exception of casein (the blocking protein used) (15, 17).Antibody specificity can be corrected by finding a new antibody, lack of blocking simply requires longer blocking periods or increased blocking solution concentration, and washing can be minimized to see resultant effect on the membrane. Each of the possible problems with the Western Blot would have to be tested by altering the procedure used above by one method (washing, antibody, blocking solution). The PCR results show templ ate replication through thermocycling, which indicates the presence of thermostable DNA polymerases in the PCR tube. From this, it can be conferred that Taq polymerase was indeed isolated. Further confirmation could be made through further purification of Taq. This could be done through 2-D SDS-PAGE vs Isoelectric point electrophoresis using the isoelectric point of Taq and using the bands emphasized as Taq, and a lower concentration gel (12). Another method would be to analyze the gel bands through other methods such as mass spectropscopy or NMR (18). There was distinct differences between three sets of Taq polymerases the standard, the sample prepared in the previous year, and the sample produced in this experiment. Most distinctly the proteins differ with respect to SDS-PAGE gels.Quite obviously, the purest of the enzymes was the standard Taq, followed by the 2011 sample, and the sample prepared in this experiment. The sample prepared through this experiment had a high amount of a salt concentration and resulted in distorted bands, along with legion(predicate) other proteins present in the sample. The enzymes also differed with respect to the Western Blot (Fig. 4). The 2011 sample failed to return 2 antibody response, indicating lack of Taq polymerase, or lack of primary antibody binding, while the standard and experimental sample both had representive banding. There may have been excessive blocking or drying of the lane containing the 2011 Taq, as the SDS-PAGE shows a representive band in the region of Taq, that is the darkest band in the lane (15). The protein concentrations as determined through the Bio-Rad assay (Tab. 1, Fig. 1) returned 7 drastically different results. The two protein concentrations differed by 2x concentration. The easiest explanation of this result is the 10x dilution was insufficient in reducing the absorbance to within the standard curve.Due to the absorbances being above the standard curve, the results are invalid, as the region in which the curve is additive is up to 0.5mg/mL (19). The 100 x dilution returned a result of 1.88 + 0.11 mg/mL. This coroborates the SDS-PAGE findings as the protein was not to a fault overloading the lane. The SDS-PAGE could have been further diluted, but the concentration used was sufficient for the purposes of the experiment. In an analysis of the PCR results (fig. 7), the brightest fluorescence bands occurred in the std., 2 and (4/Taq) lanes. This would indicate the highest activities occuring in these lanes. When compared to the western blot, the darkest banding of regions of Taq (5,?,*) returned the bands with less fluorescence. This result shows that the amount of enzyme may inhibit the PCR reaction as the the bands with the highest recoveries returned the lowest fluorescence. With an assessment of the basepair length, reaction time, and amount of enzyme used, an approximately activity of 834.5 + 63.9 bp/min/g of protein, or 3922.3 + 192.9 bp per minute. In comparison to the literature values of the protein, this is slightly above the 60 base pairs per second value, however, that was at 70 C (7).The rt-PCR returned a consistent melting temperature of 81C (Fig. 6)for all amplicon samples indicating the lack of a primer-dimer formation. Threshold was initially reached at 20 cycles (Fig. 5), which an RFU value of approximately 9000. This indicated a high activity of the taq polymerase used, at least above 1.25 Units (20). Both PCR assays agree with one another. There was no primer dimer formation say on the agarose gel, or the melt curve analysis. There was a high activity of the enzyme sample isolated as found through the bpmin-1 and cycle of reaching threshold, however, between the two assays, the rt-PCR has the significant advantage of time, and no electrophoresis required. Currently, Taq is widely available and would likely be cheaper to simply purchase commercially. This experiment does however outline a method for thermostable protein isolation which could be used for the more recent and more valuable thermostable enzymes (Pyrococcus furiosus Polymerase) which Isolation of Recombinant Taq Polymerase for PCR are superior to Taq in both thermostability, and error rate due to proofreading ability (21). Overall, the purpose of the experiment was met.Taq was indeed isolated from a culture of recombinant E. coli. This was confirmed through the Western Blotting, and thermostable DNA activity in the PCR and rt-PCR. The purity was assessed and found to be below that of the methods used by Engelke et al., 1990. The purity could be increased through use of a cation exchange column (9). The length of heat denaturation and an automatic heat controlled shaker would help to remove excess proteins and improve purity. The length of dialysis time would need to be increased for less band distortion in SDS-PAGE, and either more selective primary antibody, increased blocking or decreased washing would be required for improved Western Blotting . For further experiments, it is suggested testing the new method modifications, and or implementing recombinant Pyrococcus furiosus Polymerase.