Energy Situation and PESTLE Analysis

Question: Talk about the Energy Situation and PESTLE Analysis. Answer: Presentation Denver International Airport that is otherwise called DIA DEN is set in Denver, Colorado, United States. DIA is the greatest air terminal in the US. Alongside this, it is the 6th busiest air terminal in the US and eighteen busiest air terminals in everywhere throughout the world. Denver has the third driving residential association organize inside the country (DEN. 2017). Denver air terminal is the principle place for Frontier Airlines just as Great Lakes Airlines. It is the fourth predominant place for United Airlines. Also, it is the main air terminal in the US that has executed an ISO 14001 authorized natural administration framework to cover the total air terminal adequately. A PESTLE examination is a successful showcasing instrument that is utilized by the business associations to assess the full scale ecological components that impactsly affect the associations. Alongside this, it is additionally utilized by the business firms to follow the earth in which they are working or likewise ready to work their organizations. A PESTEL investigation is just significant to examination large scale natural factors, for example, political, financial, social, innovative, legitimate and ecological components (Wilcox and Jenkins, 2015). Denver International Airport utilizes this device to find the outside components that affect its tasks. The PESTEL investigation of Denver is given as underneath: Political Factors: Political Factors affect the associations with respect to government guidelines just as legitimate issues; and furthermore depict both formal casual principles in which business firms are obliged to work their organizations. Political strength, government approaches, exchange limitations, charge strategy, ecological guidelines, business work law, duties, etc are the significant instances of these political variables. Political elements may shift from one country to other country. Be that as it may, the air terminal industry is comprehensive and spreads the whole globe (Prather, 1998). Alongside this, concurring the report, the flying business has been created roughly $709 billion in the year 2016. The administration guidelines of the country focus on concerns, for example, quiet security, wellbeing, and some different issues. In addition, in the assembled expressed, in 1978, the deregulation happened that decreased the limitations inside the business. Deregulation canceled the principles just as guidelines those were related with the costs and section of the air terminal association into household markets. As an outcome, the prevalence of the business expanded. The expanded fame additionally upgraded the incomes of the air terminal industry in the United States. Then again, it ought to likewise be noted down that, the situation isn't the comparative in all the business sectors. It is a direct result of certain business sectors are exceptionally managed in the United States. The exceptionally managed markets lessen the incomes of the associations (Haberberg and Rieple, 2008). In addition, in such markets, the worldwide flights are exceptionally directed instead of household (national/home) flights. Likewise, Denver International Airport (DIA) additionally works its business in an intensely controlled world of politics. It is a result of in carrier industry travelers are more important than aircrafts and their security is the principle worry of the legislature. At the end of the day, it likewise can be said that, traveler security must be predominant for the carrier associations. By thinking about the patient security, the US government has set up exacting guidelines for the tasks of carrier associations (Schloh and Hardship, 1996). As a result, unmistakably, DEN works in an exceptionally controlled political environment of the country. The United States has severe guidelines for the air terminals and aircrafts. It is a result of such associations face various dangers that may reason for mishaps, irremediable harms, and passings of travelers. Thus, the US government has been made and executed heaps of arrangements, rules, and guidelines to ensure travelers in inadvertent cases. In this way, Denver needs to satisfy all the guidelines and guidelines to work its business inside the country. Then again, the legislature of the country additionally offers loads of relaxations to air terminals and carriers. The legislature is severe for the patient security not for the aircraft business of the country. It is a direct result of carrier offers various advantages that improve the economy of the country (Williams and Curtis, 2007). Therefore, the administration of the US produces an incentive by being partners of the carriers organizations. The world of politics has been loose in certain business sectors just to lead association in an efficacious and a progressively extensive way. The alliance between the administration and aircraft organizations isn't valuable for these two however advantageous for travelers moreover. Alongside this, modern guideline and world of politics in a planned way give advantages to everywhere throughout the world. In this way, it is clear that administration guidelines just as political variables of a country assume various significant jobs to choose the development and plenitude of the aircrafts organizations inside country. In this manner, clearly Denver air terminal works its business in a very managed political milieu. DIA is likewise getting a charge out of bunches of advantages because of the connection between the US government and the carrier business. The association has helped the benefit and incomes of the carrier organizations (Kerzner, 2012). In this manner, Denver International Airport is doing great so as to heighten its relationship with the legislature of the country. Financial Factors: Economic variables have various huge effects on the development and achievement of the business associations. For case, monetary development, swelling rates, financing costs, interstate charges, trade rates, embargoes, downturn, joblessness, wage rates, typical cost for basic items, credit accessibility, etc are the major monetary elements. Further, these monetary elements can be separated into two variables like: full scale financial and small scale monetary components. These the two variables accentuation on all the occasions those affect the interior just as outside monetary condition. The miniaturized scale financial occasions are identified with the practicality and inside adequacy of the business (Liebhold, Work, McCullough and Cavey, 2006). Additionally, the large scale monetary occasions contain loan costs, swelling rate, downturn, conversion standard, joblessness, working hours, financing accessibility, etc. Then again, financial elements of a country have enormous impact on the carrier organizations. The US carrier industry has seen various descending spirals. The air terminals and carriers of the country are as yet not recouped from the results of the 9/11 assaults. Alongside this, Denver air terminal is influenced by the ongoing downturn just as moderate monetary development of the country. The monetary insecurity of the country decreased the incomes of the DIA and other comparative organizations. Additionally, on account of the downturn, Denver air terminal likewise decreased its expenses to achieve upper hands over the contenders (Whitehead, 2012). Also, the drawn out downturn, oil costs changes, and an approaching worldwide log jam are other upset financial components that are influencing the development and achievement of DIA inside the economy of the country. Denver is attempting its best to oversee and hold its travelers adequately (Schloh and Hardship, 1996). As a result, it has executed a mechanized stuff framework to diminish delays, gather holding up times at baggage claims, decrease carrier work costs, and improve the notoriety in the worldwide market. DIA is additionally managing expanded high fuel costs and working upkeep costs in an adequate way. Aside from this, the downturn had abandoned various beneficial outcomes it. Because of the downturn, air terminals like Denver air terminal have changed their style of working. They likewise improved their administration principles and furthermore discounted their costs to draw in an ever increasing number of clients. The air terminal is a base of the entire aircraft indust ry. In current, the business is altering its business according to the changing monetary states of the country (Zalengera, Blanchard, Eames, Juma, Chitawo and Gondwe, 2014). Be that as it may, a great deal of development is assessed in the up and coming years; and because of the development of aircraft industry, the eventual fate of Denver air terminal can be seen splendid later on timespan. Social Factors: Social variables or socio-social elements investigate the social condition of the market, and measure determinants, for example, socioeconomics, social patterns, populace examination, etc. Alongside this, these components include the common conviction just as mentalities of the masses. These socio-social variables contain populace development, social angles, social desires, wellbeing cognizance, populace elements, age appropriation, vocation perspectives, standards, an Earth-wide temperature boost, and so forth. These social elements center around the security and wellbeing of the masses (Ho, 2014). What's more, high patterns in socio-social variables affect the interest and flexibly for the items or administrations of the organizations. Business associations may change their administration methodologies to take focal points of these social patterns for the development and achievement of the business. Then again, it is notable that Denver International Airport is the greatest air terminal in the United States. It thinks about social variables to improve the matter of the carrier business. Additionally, air terminals and carriers create social qualities for the development and achievement of the organizations. There are bunches of enterprises that gain benefits from the carriers (Wilcox and Jenkins, 2015). Air terminals and aircrafts are significant methods for transportation for the individuals and business associations. The two of them are profoundly affected by the social factors and powers too. In this circumstance, the DIA has scaled down its costs to address the issues and requir

A Guide to Ethnomethodology

A Guide to Ethnomethodology Ethnomethodology is the investigation of how individuals utilize social collaboration to keep up a progressing feeling of reality in a circumstance. To assemble information, ethnomethodologists depend on ​conversation examination and a thorough arrangement of procedures for efficiently watching and recording what happens when individuals interface in characteristic settings. It is an endeavor to arrange the moves individuals make when they are acting in groups.â Causes of Ethnomethodology Harold Garfinkel initially concocted the thought for ethnomethodology at jury obligation. He needed to clarify how the individuals sorted out themselves into a jury. He was keen on how individuals act specifically social circumstances, particularly ones outside of the every day standard like filling in as a juror.â Instances of Ethnomethodology A discussion is a social procedure that requires certain things with the goal for members to recognize it as a discussion and prop it up. Individuals take a gander at one another, gesture their heads in understanding, ask and react to inquiries, and so forth. In the event that these techniques are not utilized accurately, the discussion separates and is supplanted by another kind of social circumstance.

The History of the Thematic Analysis Psychology Essay

The History of the Thematic Analysis Psychology Essay Thematic analysis is an analytical technique serving to interpret the contents of texts or human experiences and widely used in qualitative research. In its interpretation of qualitative data, thematic analysis generates themes within the distinguished categories of data. Each category that reflects a recurring feature of the obtained data attributes a code to a corresponding informational block. However, data categorization is thematic analysis pursues the aim of managing the overall volume of unstructured data rather than indicating conceptual issues. In this vein, thematic analysis entails a clear and precise definition of codes that are distinct from one another. During analytical procedures, a researcher refers to the developed code log book to categorize data and allow themes to emerge.[1] In other words, a theme in thematic analysis represents a statement of meaning observed in most of the data. This meaning may be also reported by a minority of participants, but carry a strong emotional power or factual impact on the studied situation. In thematic analysis, themes may be regarded as units formed by deriving and comprising patterns of conversation issues, meanings, feelings, recurring activities. Proverbs, and vocabulary. As such, themes serve to bring together elements ideas, perceptions, or experiences that may seem meaningless when considered separately.[2] In other words, the role of thematic analysis is to summarize the data content and draw relationships between distinct codes to produce a social meaning shared by the examined population sample. Therefore, thematic analysis is an interpretive analytical technique dedicated to revealing new concepts and ideas. Thematic analysis is theoretically flexible and allows answering various research questions without adherence to a specific framework. This analytical approach is mostly beneficial for responding to research questions regarding human experiences, ideas, attitudes, and perceptions.[3] Its focus on motifs and beliefs driving individual behavior fits scholar ship in psychology, social studies, behavioral research, and more applied disciplines like education, health, and clinical research. Psychology is a discipline that seeks to explore and analyze human experiences and behaviors to determine factors and personality traits shaping individual’s behavior patterns. In this vein, qualitative studies represent a significant portion of psychology scholarship when researchers attribute meanings to different behavior types observed or discussed with human participants.[4] In line with the prevailing qualitative research design, psychological studies focus on synthesizing the attained data and interpreting results into meaningful conclusions. In this pursuit, scholars tend to use different names to describe content analysis procedures utilized in their inquiries or using them without a specific description. Regardless of the expressed position, psychologists appreciate the thematic analysis framework for several benefits offered. While examining human observations, feelings, and attitudes, this analytical approach tends to bridge the researcher and participants intellectuall y and emotionally, fostering understanding of the former of motifs and beliefs underpinning behavior of the latter.[5] Moreover, thematic analysis allows combining different research methods that is a substantial advantage for psychological research. For instance, thematic analysis is applicable to processing and interpreting data obtained through both qualitative and quantitative methods. In thematic analysis, interview findings may be complemented by survey results to expand the depth and breadth of the acquired understanding while strengthening validity and reliability of the produced conclusions.[6] This argument is crucial in terms of qualitative research criticism that stresses on high subjectivity and descriptiveness of qualitative research findings. When applied to a single research method, thematic analysis serves to organize the unstructured dataset and embed a systematic approach in the analytical process. In the case of psychological research utilizing several research methods, thematic analysis is useful to generate an insight into the studied phenomenon while proving credibility of the attributed meaning. References Boyatzis, Richard E. Transforming Qualitative Information: Thematic Analysis and Code Development. Thousand oaks, CA: Sage, 1998. Gibson, Will. “Qualitative Research Analysis in Education.” In Educational Research and Inquiry: Qualitative and Quantitative Approaches, ed. Dimitra Hartas, 295-307. London: Continuum International Publishing Group, 2010. Goodwin, C. James and Kerri A. Goodwin. Research in Psychology: Methods and Design. Hoboken, NJ: John Wiley Sons, 2016. Thatchenkery, Tojo and Carol Metzker. Appreciative Intelligence: Seeing the Mighty Oak in the Acorn. San Francisco, CA: Berrett-Koehler Publishers, Inc., 2006. [1] Ibid. [2] Tojo Thatchenkery and Carol Metzker, Appreciative Intelligence: Seeing the Mighty Oak in the Acorn (San Francisco, CA: Berrett-Koehler Publishers, Inc., 2006), 235. [3] Ibid. [4] C. James Goodwin and Kerri A. Goodwin, Research in Psychology: Methods and Design (Hoboken, NJ: John Wiley Sons, 2016), 302. [5] Richard E. Boyatzis, Transforming Qualitative Information: Thematic Analysis and Code Development (Thousand oaks, CA: Sage, 1998), 6. [6] Goodwin and Goodwin, 302.

Capital Punishment Should Be Abolished - 1137 Words

The case for capital punishment Topic: why capital punishment should continue General purpose: To argue Specific purpose: my main aim is to convince the audience that capital punishment should be upheld. In other, the advantages of capital punishment outweigh its disadvantages. Introduction Capital punishment is also known as death penalty and it has been in existence since time immemorial. Throughout history, the death penalty has been used to punish a number of crimes that include murder and other premeditated crimes. However, there has been growing campaigns calling for the abolishment of capital punishment. Many of these campaigns have been spearheaded by religious groups and other interested parties. As a Christian, I am of the opinion that such arguments have no biblical bases because the bible advocates for the application of the death penalty for certain crimes like witchcraft, murder and kidnapping. Whenever the word capital punishment is mentioned, it raises a lot of heat from all the sides of extremist. One side argues that capital punishment should be abolished because it may result in the execution of an innocent person while the other side arguers that capital punishment deters crime. As noted earlier, the death penalty is one of the most discussed topics in the criminal justice system. The death penalty used to be legal in the united states until 1972 when the supreme court in Furman v. Georgia ruled it unconstitutional arguing that it violatedShow MoreRelatedCapital Punishment Should Be Abolished965 Words   |  4 PagesHaesemeyer Advanced Studies English 9 7 April 2017 Capital Punishment Over the centuries, capital punishment has fallen in and out of public support. In several countries, the practice has been overruled by law. In others, it is simply not exercised. More than half of U.S. states still practice capital punishment for capital crimes. Often, innocent people are sentenced to death because of circumstantial evidence. Capital punishment should be abolished in all fifty U.S. states because of the severalRead MoreCapital Punishment Should Not Be Abolished1541 Words   |  7 Pagestime, if a person committed a severe crime, like murder or rape, they were executed to maintain peace in the community and to bring comfort to those who knew the victim. Capital punishment has been used in almost every part of the world, but in the last few decades many countries have abolished it. The issue of capital punishment has been a sensitive topic for nations attempting a careful balancing act between prisoner’s rights and legal defense teams and society’s la ws on cases of extreme gravityRead MoreCapital Punishment Should Not Be Abolished901 Words   |  4 Pagesgovernment (â€Å"Capital Punishment in the United States†). Capital punishment is being debated all over the world whether it is murder or justice for the crime they have committed. Statistics show that murderers often kill again after releasement from prison. The Bureau of Justice gives relevant statistics pertaining to murderers who were released from prison: in 1994 40.7% of murderers were arrested for a new crime within three years of release (United States Department of Justice). Capital punishment shouldRead MoreCapital Punishment Should Be Abolished1115 Words   |  5 Pagesjail time, house arrest, and/or having to pay fines. Crimes that are severe can lead to greater punishment, like spending life in prison. The most severe crimes can lead one to an equally severe punishment known as capital punishment. Capital punishment is the authorization to kill someone fo r the crime he or she has committed. Capital punishment, commonly referred to as the death penalty, should be abolished in all states because it can put innocent lives at risk, it costs millions of dollars each yearRead MoreCapital Punishment Should Not Be Abolished750 Words   |  3 PagesCapital Punishment Should Not be Abolished There are many reasons why the United States of America keeps capital punishment. These reasons include the deterrence theory, the idea of retribution, cost of prisons, and general safety of the public. First of all, I’m sure that you have heard of the deterrence theory. Deterrence is basically the fear of punishment; and even though it doesn’t prevent all crimes, the results are still undeniable. The deterrence theory is when criminals thinkRead MoreCapital Punishment Should be Abolished883 Words   |  4 Pages Capital punishment is the punishment of death for a crime given by the state. It is used for a variety of crimes such as murder, drug trafficking and treason. Many countries also have the death penalty for sexual crimes such as rape, incest and adultery. The lethal injection, the electric chair, hanging and stoning are all methods of execution used throughout the world. Capital punishment has been around since ancient times; it was used in ancient Rome, and one of the most famous people to be crucifiedRead MoreCapital Punishment Should Be Abolished1955 Words   |  8 Pagesit has become apparent that capital punishment should be abolished worldwide. Around t he world, various countries continue to practice this brash, inhumane punishment. The legal system is intended to regulate citizen’s behaviour which aims to provide a smooth functioning society. When someone is sentenced to death, the legal system loses the â€Å"fair† aspect of the proceedings, as under no circumstances is the death penalty â€Å"fair†. Capital punishment should be abolished because it violates Section 12Read MoreCapital Punishment Should Be Abolished1135 Words   |  5 Pagesname -unknown. Nobody should ever have to hear these words. Especially coming from their own state the one that is supposed to protect their rights no matter what. Capital punishment should be abolished from the United States. The number one goal of the constitution is to protect the life of an individual and the death penalty passes by this. The death penalty is an easy way out of the crime, it is expensive, philosophers and even psychologists disagree with capital puni shment and most important itRead MoreCapital Punishment Should Be Abolished Essay1293 Words   |  6 PagesApproaching the topic of capital punishment is difficult when looking at society as a whole. In the opinion stated by my class group, capital punishment should be entirely abolished due to the possibility of mistakes while sentencing. This idea relies on the basis that capital punishment has absolutely no purpose or benefit. Members of my group mistakenly used a utilitarianist point of view to argue their case, even though this view can be stretched to fit almost any argument. This paper will exploreRead MoreCapital Punishment Should Be Abolished1439 Words   |  6 Pagesto outlaw capital punishment in America completely. We are meant to be better and bigger than this then why do w still have this barbaric law in practice it is inhumane, morally wrong and expensive. In my paper I will try to cover the taboo topic of capital punishment and why it should be abolished because if we as a society want to grow and advance in a sophisticated society we must abolish the death penalty all across the land. To begin with I must explain what capital punishment really is and

Find Out Whos Concerned About University of Chicago College Essay Samples and Why You Should Listen to Them

Find Out Who's Concerned About University of Chicago College Essay Samples and Why You Should Listen to Them Up in Arms About University of Chicago College Essay Samples? There's, clearly, a limit on the range of pages even our very best writers can produce with a pressing deadline, but usually, we figure out how to satisfy all the clients seeking urgent assistance. Furthermore, the high quality and originality of the item is guaranteed. At times, the best method to learn and understand new information is by way of seeing and understanding work which is already completed. You don't have to be worried about your personal details that could be viewed, as we handle the matter on a safe network. I have always enjoyed riding in cars. Explain the auto connection better. Descriptive essay about an auto accident. The exact same is true in the United States of america. Thus, State University isn't only the ideal location for me, it is the sole spot for me. On top of its growing cultural and ethnic diversity, it is becoming a master at creating a niche for every student. The University of Chicago is known for quirky and elaborate essay prompts. Who Else Wants to Learn About University of Chicago College Essay Samples? Last, don't neglect to connect your very own past experiences and accomplishments to the resources you decide to highlight. Student loans aren't free money and has to be paid back. The findings reveal that in Malaysia, it's quite complex to demonstrate negligence in instances of road accidents because proving a case is contingent on the elements to be proven. Yet, three decades later, not one of that transpired. The application text needs to be effective to put in the college. All you need to do is to locate reliable sources of information which offer writing tips. You have to construct a good outline before you write. Inside this section you'll also find more comprehensive info about each stage in the application approach. 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GFP protein Free Essays

string(97) " the low salt beaker and in the column chromatography, causing GFP to spread down the container\." Green Florescent Protein, abbreviated as GFP, is a protein composed of 238 amino acids that is commonly found in mnemiopsis, comb Jelly. It has a major wavelength at 396 nm and a minor one at 475 nm. GFP is what gives mnemiopsis their bright green florescent glow. We will write a custom essay sample on GFP protein or any similar topic only for you Order Now ultraviolet light, or blue light, is necessary to see the florescent glow of this protein. GFP is an irregular protein because It Is highly resistant to denaturation by temperature and PH. It can survive In temperatures up to 98 degrees and has a pH of 12. 2 due to Its complex exterior, called the beta barrel. At an pH higher than 12. It denatures. It also has an Isoelectric point at 5. 3. The peripheral beta barrel cannot be digested or broken apart by protease because of the strong bonds holding It together. The beta barrel protects the chromophore, which Is the substance which gives GFP Its green glow. When CFP Is extracted from the plasmid of an E. Coll or from a Jellyfish, It contains an array different contaminants making it difficult for scientist to do experiments with GFP. A procedure in purifying GFP from a crude cell extract to nearly 100% GFP so that it can be analyzed and used in scientific experiments and research is necessary. The goal is to ptimize each protocol used to purify crude GFP. Methods Ammonium Sulfate Precipitation To purify the crude samples of GFP, the ion exchange method separates substances inside the test tube by similar charge. A sample of crude GFP of 7. 5 mL in a plastic tube was used for the experiment. Knowing that 43. 6 grams of ammonium sulfate in a 100 mL solution yields a 70% percent saturated solution, the proportion 43. 6g 11 00 mL=x/7. 5 mL was used to determine that 3. 27 grams of ammonium sulfate needs to be added to the experimental sample. After adding the ammonium sulfate, the solution was stirred gently to prevent frothing. Once most of the solution is transferred, the tube was placed on a triple beam balance along with another tube that went through the same process. The centrifuge was set at 15,000 rpms for 15 minutes so that the hydrophobic materials will separate and become the supernatant while the GFP pellet will remain behind. Once the 15 minutes elapsed, a new pipette was used to remove the supernatant, leaving behind the pellet of GFP and hydrophilic contaminates. To remove the hydrophilic substances, 5 mL of 4 molar ammonium sulfate and 15 mL of 10 mL tris at a p of 8 was added Into the oak ridge entrifuge test tube. The solution Is then put Into the centrifuge at 15,000 rpm for 15 minutes again. Once 15 minutes has passed, the supernatant, containing the GFP, was removed by a pipette and put In a microfuge. Hydrophobic Interaction Hgure yaropnoDlc Interactlon set up One molar ammonium sulfate was added to the column to wash the sample. Adding 1 molar ammonium sulfate washes the sample because a high salt concentration increases hydrophobicity of the GFP and the buffer, causing most of the GFP to be at the very top of the column. Substances that are hydrophilic get flushed out of the olumn while the more hydrophobic substances stay in the container. After the column has been eluted with 1 molar ammonium sulfate, the tris buffer is added to the ammonium sulfate to dilute it into . 5 molar ammonium sulfate. The volume of 1 molar ammonium sulfate inside the oak ridge centrifuge test tube is the volume of the tris buffer that will be added. After the column chromatography has been flushed with . 5 molar ammonium sulfate, more hydrophobic substances will be flushed out since the hydrophobicity of the tris buffer and the GFP has decreased. This causes the GFP to spread out in the column. Finally the amount of . 5 molar ammonium sulfate is diluted with tris buffer to . 5 molar ammonium which should cause most of the GFP to be flushed out of the column along with other substances that are very hydrophobic. While this experiment is going on the liquid that comes out of the column is collected in multiple test tubes. These test tubes contain GFP and other contaminants. The solutions are than read by a spectrophotometer. Each test tube wil l be tested by the spectrophotometer so that a graph can be made. Anion Exchange Figure 2: Siphon Bridge set up for Anion Exchange Figure 3: Centricon Test Tube In order to use anion exchange, the starting condition of the sample needs to be in a low salt solution. However after the GFP had gone through hydrophobic interaction, it was in a high salt solution. Before facing this dilemma, the fractions were pooled by centricon which decreases the overall sample volume by removing some buffer and salt solution. This greatly increases the GFP concentration in the samples. The fractions are placed in the centricon and then into a centrifuge for 25 minutes at 3,000 rpm to be separated by size. The large proteins stay in the entricon while buffer and salt solution goes into the plunger. To reduce the concentration of salt in the GFP sample, the sample is diluted 10 folds. Since the amount of GFP that was restored was 18 mL, 162 mL of tris buffer needed to be added. The diluted GFP is then put in the chromatography container, containing positively charged DEAE which is attracted to the GFP at a low salt concentration. After the GFP has been completely filled, the column is connected to a beaker that contains a low salt concentration. the low salt concentration beaker is connected to a high salt concentration beaker. As one drop of low salt solution goes into the chromatography column, one drop of high salt solution goes into the low salt solution. Gradually the salt concentration increases in the low salt beaker and in the column chromatography, causing GFP to spread down the container. You read "GFP protein" in category "Papers" The eluted GFP dripped out of the column chromatography to be collected in test tubes. I nree pnase partltlonlng Figure 4: Precipitate of GFP. T-butanol is one top while contaminates are on bottom GFP then went through three-phase partitioning, also known as TTP. The fractions taken after an anion exchange was 15 millilieter. Ten ml of 4 M ammonium sulfate was added to this volume to increase the salt concentration of the solution to 1. M, which is about 40% salt saturation. Twenty-five milliliters of t-butanol was added then added which was the same amount of ammonium sulfate and GFP in the container. The container was then placed in the centrifuge for ten minutes at 4600 RPM, causing the mixture to split into three layers; butanol on top, GFP in solution on the bottom, and precipitated contaminants in-between. The top layer of butanol and disk of precipitate were taken out. The volume of GFP solution was again matched in utanol and the container went into the centrifuge again. An aspirator was used to extract the GFP into a microfuge. . 6M ammonium sulfate was then added to the microfuge and the container was placed in a micro centrifuge for one minute at 13,000 RPM. Butanol and other contaminants that had not been take out previously formed a disc, was then taken out with an aspirator and the remaining GFP was then left in the microfuge. HPLC Figure 5: HPLC basic layout After the sample went through three phase partitioning, it was put through the High Performance Liquid Chromatography for a final purification. First liquid was put into the HPLC to clean out any previous GFP inside the loop of the HPLC and the column of the HPLC. Then, GFP in the microfuge was sucked into an injector to be put into the HPLC. Pushing the top of the injector slowly, GFP entered into a loop inside the HPLC. Once the GFP was placed in the loop, a knob was turned clockwise to the word lock. The GFP was then sent to the column where it was purified further by size through the minuscule beads. About 6,000 pounds of pressure per square inch was produced by the HPLC to push the GFP through the beads. While this was occurring, a pectrophotometer connected to the HPLC read the wavelengths of substances. Near the 396 nm wavelength, GFP was collected in a microfuge tube. A UV light was held near tne exlt 0T Results e HPLC to measure tne amount ng sample. Graph 1: Results of the sample after HIC at a wavelength of 395 nm Graph 2: Results of the sample after HIC at a wavelength of 280 nm Graph 3: Results of the sample after HIC of the entire spectrum Seventeen test tubes were received after the HIC purification process. A blank consisting of tris buffer and ammonium sulfate was sampled in the spectrophotometer against liquid from each of the seventeen test tubes. Graph one represents the sample after HIC at a wavelength of 395 nm while graph two Results shows the results after HIC at a wavelength of 280 nm. After HIC, the fractions 12 to 16 were chosen for their purity and recovery of GFP. Graph one show the amount of GFP in each fraction number while graph two shows the total amount of protein in each fraction number. Graph three shows the spectrum of the entire sample. Graph 4: Results after Anion Exchange at a 397 nm wavelength Graph 5: Results after Anion Exchange at a 280 nm wavelength Graphing 6 Thirteen test tubes were collected from the Anion Exchange purification process. This time the samples were blanked against tris buffer at 8. 0 pH and 0. 5 molar sodium chloride. Graph four shows results of the Anion Exchange at a 397 nm wavelength and graph five shows the results after Anion Exchange at a 280 nm wavelength. Once again, the graph at a 297 nm wavelength shows the amount of GFP while the graph at a 280 nm wavelength shows the amount of total protein. Graph six represents the results of the entire spectrum. The GFP peak was a lot more visible. Step Iotal sample (mL Abs (280) Total Protein Abs (397) GFP Ratio Crude sample 120 1600 . 25 At-ns042- 20 1 . 61 . 9 118 HIC 18 . 28 . 173 . 618 15 . 126 . 130 1. 03 3 Phase Partitioning . 01 n/a . 75 . 243 . 257 1. 06 Table 1: This is the overall data table. The second column shows the total volume at the start of each purification step. The following two columns are the peaks of the graphs at those wavelengths. The last column represents the ratio of GFP to the total Protein. The most desirable ratio is 1. 2 5. Dlscusslon The first method in purifying the crude GFP was using the ammonium sulfate precipitate. When ammonium sulfate is placed in water, it dissociates into ammonium (NH4+) and sulfate ions (S042-). Water, composed of two hydrogen ions and one oxygen ion, is a polar molecule because the oxygen has a high electronegativity. Oxygen has a greater affinity making the oxygen portion of water negative and the hydrogen portion of the water positive. The dissociated positively charged ammonium ion is allured to the negatively charged oxygen while the negatively charged sulfate ions are attracted to the dissociated positively charged hydrogen. The attraction between the ammonium sulfate and the water was so strong that the GFP and other proteins were left unoccupied, causing them to precipitate. When GFP in the 70% salt solution was placed into the centrifuge, substances such as DNA and RNA was removed because they became part of the supernatant. At a 70% salt concentration, only hydrophilic substances stay in solution while the more hydrophobic substances precipitate. When the GFP in a 25% solution of salt was placed in the centrifuge, the GFP and other substances went back into solution because there not enough water was occupied by the salt. Before the GFP is placed in the centrifuge, it must be balanced with another centrifuge with the same weight and the two containers must be placed across from one another. This is vital because the centrifuge needs to be balanced when it is rotating at an incredibly fast speed. Failure to have balanced centrifuge containers can result in a broken centrifuge and loud sounds. Also when mixing the GFP with salt, it is important not the shake the container or frothing will occur, making it difficult to transfer the solution in to an oak ridge centrifuge tube. The second purification procedure that GFP underwent was hydrophobic interactions. During this purification, GFP binded to the non-polar Phenyl Sepharose beads because of its non-polar and hydrophobic traits. However the water in tris buffer is strong enough to separate the attraction between GFP and the Phenyl Sepharose. Therefore a high salt concentration is necessary to occupy the water so that the GFP and the Phenyl Sepharose to be attracted together. At a high salt concentration, GFP with bind easily to the Phenyl Sepharose since very little water molecules would interfere with the attraction and at a low salt concentration, GFP would not bind easily to the Phenyl Sepharose because tnere wlll De a lot 0T unoccuplea water molecules tnat wlll De aDle to InterTere wltn the GFP and Phenyl Sepharose attraction. Before the experiment, ten millimolar tris buffer at a pH of 8 was used to clean the column in order to keep the pH stable and to wash away the salt, ammonium sulfate, in the column. Removing the salt is vital because the buffer that once surrounds the salt will be allured to the hydrophobic benzene and to the hydrophobic patches on the GFP. Since the hydrophobic patches of the GFP are already filled, they will be flushed out, leaving mostly beads of benzene and the 10 millimolar tris buffer at a pH of 8. Once the column has been clean, it needs to be equilibrated so that the salt concentration is the same through the olumn. The step gradient used, started ata 1 molar ammonium sulfate concentration and was halved until a . 25 molar concentration to separate substances by hydrophobicity. The third purification procedure was anion exchange. In this procedure, GFP and other contaminants are separated by charge. The beads in the containers are different from the beads from the hydrophobic interaction because on they have a different chemical called DEAE which makes them positively charged. GFP has both protons and electrons on it which is why it was not easily attracted to the DEAE, which is why the GFP is put in a basic solution. Ata high pH, the amount of negatively charged hydroxide increases and these hydroxides are allured by the protons on the GFP. The protons are than neutralized, making GFP a negatively charged molecule. The isoelectric point of GFP is at a pH of 5. 3. Ata pH higher than 5. 3, it is negatively charged and when it is at a pH lower than 5. 3, it is positively charged. Once the column chromatography is filled with GFP and connected to a beaker of low salt which connected to a beaker of high salt, anion exchange occurs. As the salt concentration increases, the GFP slowly spreads down the column and eventually out f the column into test tubes. Between the HIC and the Ion exchange chromatography, the sample the fractions were pooled and put in a centricon causing the GFP concentration in the samples to increase. This occurred because the ultrafilter only allowed particles smaller than protein to go in to the pusher. The large proteins stay in the centricon while buffer and salt solution goes into the plunger. The sample of GFP was also diluted 10 folds because the sample needs to be in a low salt solution to use anion exchange and after the GFP had gone through hydrophobic interaction, it was in a high salt solution. The anion exchange method creates a continuous salt gradient because as one drop of low salt solution goes into the column chromatography, causing GFP to spread down the container. The follow procedure was the three phase partitioning purification. T-butanol and 1. 6 molar ammonium sulfate were essential for this procedure. T-butanol has a low density causing in to stay above the GFP solution. In addition it has an attraction for water and other hydrophobic substances causing 5 mL of water to be drawn out of the GFP sample and precipitated substances to float between the t-butanol and the GFP sample. Fresh t-butanol is necessary after removing the old t-butanol with the contaminants because at that point, the salt concentration had increased since water was drawn out. was aDle to De preclpltatea Decause 0T tne nlgn salt concentration. The final procedure for purifying GFP was using the HPLC which separated substances by size. The beads used in the HPLC column are miniscule and porous. The pours on the beads give substances of the same size more opportunities to leave the HPLC at the same time. Since the beads are so small, high pressure is needed to push the GFP sample through the beads. Naturally, smaller substances will exit the HPLC first while larger materials will exist last. In all scientific experiments room for error is unavoidable. During the HIC, IEX, three phase partitioning, and the HPLC, amounts of GFP were lost due to the GFP sticking to a container, a pipette, and even spills. During the HIC some of the GFP was lost due the overflowing the test tubes with liquid exiting the column. During the HPLC some GFP was lost because not all GFP dripping out of the HPLC went in to the microphage. Other errors include letting the column dry because the liquid was not dded to the beaker about the column. During the spectrophotometer runs, the blank was no inserted correctly causing the reading of the GFP to be incorrect. In addition, the order in which the GFP samples were suppose to be placed in the spectrophotometer was messed up. Judging from the overall purification table, table 1, the purification was quite successful. Originally, the ratio was only . 25, but by the end of all the purification procedures, it obtained a ratio 1. 06. A 1. 25 ratio is most desirable and through the purification, the ratio was nearly reached. The anion exchange, three phase artitioning, and the HPLC purification were the most impacting procedures. The anion exchange greatly increased the purity of the crude sample compared to the HIC purification. The three phase partitioning and HPLC purified the GFP even more. Some improvements to the protocols would be to start with the anion exchange purification so that overall, the salt solution would go from a low salt concentration to a higher salt concentration. This also eliminates the need to dilute the solution. In addition, an automatic machine could be used to shift the test tubes that collect the iquid exiting the columns to prevent overflowing test tubes and the risk losing GFP. GFP is unique because of its florescent glow. This glow can be used as a marker or an indicator. If a glowing marker could be placed on infectious cells such as tumor cells or cancerous cells, it would revolutionize the treatment of these diseases because doctors will be able to track where the harmful cells are. In addition, if it is possible to trigger the florescence of GFP with UV light, it can eventually be used in light bulbs to produce light. GFP light bulbs would last for an incredibly long time ince they are very resistant to denaturing. In addition, in vehicles, GFP can be mixed in the motor oil, transmission oil, power steering oil, air conditioning oil, and other oils so that if a leak occurs in a car, it can easily be spotted by shinning UV light on the car. The purification of GFP can lead to endless new innovations in electrical engineering, automotive repair, and curing deadly diseases. How to cite GFP protein, Papers

The Business Network free essay sample

How are these two approaches different? Does it depend on the state of the economy which one should be used? Why? Before the dot-com debacle Cisco Systems used â€Å"push† approach – a lot of inventory was made, and it was based on best-guess forecasts. This approach was not bad when economy was strong and there was huge demand and there was no need to predict precisely the inventory. When during the recession of the economy Cisco Systems implemented â€Å"pull† approach, which rely on precisely planning of inventory. The second approach allows having free costs because of reducing inventory and also freeing up working capital. 2. What are the different elements that need to come together to bring supply chains to the optimal levels needed by these companies? What role does IT play? In order to bring supply chains to the optimal level company needs to coordinate supply chain operation system with IT technologies. We will write a custom essay sample on The Business Network or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page Also, in order to work in appropriate manner system needs to get and adapt information from marketing, finance, sales and supply department. IT plays a very important role: its connect the whole system, coordinate it and adapt. . How are the approaches to inventory management taken by O’Reilly Auto Parts, on one hand, and Cisco Systems and Black amp; Decker, on the other, different? O’Reilly Auto Parts uses inventory as a competitive differentiator and try to increase customer service levels and replenish inventory on a nightly basis, while at the same time managing an increasing number of products. Also its strategy relies on partnership between the supply chain operation and IT. Black amp; Decker chose another strategy: they try to lower inventory levels to free up working capital.