Plasmids Lab Report Example | Topics and Well Written Essays - 5500 words

Plasmids - Lab Report Example Introduction: Plasmids are extrachromosomal DNA present in the bacterial species. They are double stranded DNA which forms circles with size ranging from 1 kb to 200kb ( kilobase ). Plasmids are very advantageous for the genetic engineering. Plasmids code for many antibiotic regions and they have the ability to accept the gene of interest. The transformation of our gene of interest into the plasmid is called recombination and the bacteria are called recombinant bacteria. Thus plasmids can be used as cloning vehicles or vectors. The first step of transformation is the isolation ofhte plasmid DNA from the given bacteria culture. The basic method or DNA isolation is 1. Cutluring the host cell containing the Plasmid DNA. 2. Harvesting and lysing the cell to separate the DNA from the cell organelles. 3. Separation of chromosomal DNA and Plasmid DNA through precipitation method. 4. Plasmid DNA isolation and purification. Since both chromosomal and plasmid DNA will remain in the solution, the method to isolate plasmid DNA from the Chromosomal DNA is precipitation method. Larger DNA molecules (i.e. chromosomal DNA), bound to the proteins are separated from the Plasmid DNA when the protein is precipitated. The plasmid DNA which remains in the solution is then precipitated using ethanol. Method: Method: 1. A single colony of Bacteria containing the pBlueSkript KS II was grown overnight in the Luria Betroth overnight with ampicillin as the antibiotic. 2. From the overnight culture, 1.5 ml of the culture was taken in the centrifuge tube and centrifuged at maximum speed for 1 minute. 3. The supernatant containing the medium is discarded and the cell pellet was kept as dry as possible. 4. The cells were resuspended in the 100Â µl of GTE buffer and mixed gently using the pipette to ensure that no cell pellets remain in the solution. 5. To the cell pellets, 200 Â µl of cell lysis buffer was added at room temperature. The tube was mixed gently by inverting the tube up and down five times and incubated at ice for 5 minutes. 6. To the mixture 150 Â µl neutralization buffer was added and again inverted gently up and down 5-6 times. 7. The mixture was centrifuged at maximum speed for 10 minutes and the supernatant was added to the new tube. 8. To the supernatant, 1000 Â µl of 100% ethanol was added to precipitate the DNA. 9. The tube is centrifuged for 10 minutes in maximum speed. 10. The supernatant was removed from the tube and to the whitish DNA pellet, 1ml of 70% ethanol was added and the tube was inverted several times and centrifuged at maximum speed for 2 minutes. 11. The supernatant was removed from the solution and to the DNA, 500 Â µl of 70% ethanol was added as final wash. The tube was again centrifuged at top speed for 2 minutes and the DNA pellet was obtained. 12. The pellet was resuspended in 40 Â µl of 10mM Tris- HCl with RNase. The tube was mixed by flicking the tube and incubated at 37Â ° C for 5 minutes. 13. 5 Â µl of the Plasmid DNA was transferred to sterile microfuge tube and was labeled as B3- 5 Â µl PKS II- southern blot and stored at -20Â °C. Result and discussion: The DNA was extracted from the culture using the miniprep method. The plasmid DNA obtained in this method is used for the transformation process. Answer 1: Ampicillin is an antibiotic that resists the growth of the ampicillin senstitive strains when added to the medium. As our plasmid PKS II codes for ampicillin gene, ampicillin was induced in the growth medium to avoid contaminants. Answer 2: RNase is the enzyme that cleaves the RNA present in the given sample. RNA are the contaminants seen along with the plasmid DNA. Hence RNase was added to cleave the RNA. Answer 3: We can use alkaline lysis/ phenolic extraction method or alkaline lysis/PEG

Art Assignment Example | Topics and Well Written Essays - 1250 words

Art - Assignment Example On the other hand, Rome is based on aristocratic and hierarchical form of government and society. The two types of pottery are hand-built and wheel-thrown pottery. There are 3 techniques in making hand-built potter namely pinch pot, coil pot, and slab pot techniques. Pinch pot technique is undertaken by kneading the clay and then shaping it into different potteries such as bowl, dish, cup, etc. The coil pot technique is achieved by making long, round clips of clay and then these are coiled and stacked to achieve the desired shape. The third technique is the slap pot which is used to make clay boxes and other geometric forms and shapes. Advanced or complicated shapes can be achieved through the combination of the techniques. The other type of pottery is the wheel-thrown pottery which uses a potter’s wheel to be able to form shapes. The use of the said tool is a needed skill thus it is the primary requirement in using this method. Another important skill is to know the amount of water to use in making the pottery. This water suspension is referred to as the slip. The potter y glaze seals and waterproofs the pottery and undertaken after the first firing process that hardens the clay. Another term important in pottery is leather-hard which means a stage wherein the pottery is considerably hard but still workable. This stage is included in the greenware stage of pottery process which is prior to being put in the kiln or the baking apparatus. Pottery at this stage is also called bisque. The Egyptian and Mesopotamian civilizations are included in the earliest human civilizations with very distinct culture and art forms. The Mesopotamian civilization originated near the Tigris-Euphrates River while the Egyptian civilization originated near the Nile River. They both have a society based on religious beliefs. This can clearly be seen in their art wherein the

Pupil Z Individual Observation Program Essay Example | Topics and Well Written Essays - 1250 words

Pupil Z Individual Observation Program - Essay Example I decided I should then carry out the two-week observation programme. For one week, I have observed Pupil Z accessing school activities and leisure time in the supportive sitting position, stimulated by his hands while exploring items on his tray. In the second week, I prioritized opportunities for Pupil Z to access school activities and leisure time by lying on his back (supine position) and exploring his surroundings with his feet. These activities were a precursor to other motor skills (hands exploration), seeking to establish the impact these activities may have on his alertness, motivation and daily learning. Settings Pupil Z’s two-week individual observation programme started when he entered the school and it stopped when he left (9.30-3.30) each session / activity. Each session lasted about 25 minutes. Pupil Z’s am/pm snacks (2 x 30 minutes) included a 5 minute break, lunch (60 minutes plus 5-minute break), and 5-minute breaks between the sessions / activities. T hese breaks were not included in the total number of minutes of observation per day (200 minutes) and 1000 minutes per week. Every morning, Pupil Z had one-to-one interactions with an adult (massage). He was observed participating in class greetings, one additional group activities (e.g. sensory story, foot spa, messy play, etc.), and a self-occupation time (sensory exploration). An interval of 5 minutes was often allowed between the 25 minutes activity / sessions in order to prevent broadness and ensure that adjustments were made to fit his energy level. Every afternoon started with a self-occupation time (Little Room), followed by a sensory exploration. Next came a group activity, and the session usually ended with the whole class saying goodbye to each other. An interval of 5 minutes was allowed between activity/session. I felt it was important to provide Pupil Z with various short, repetitive, and well-cued routines (e.g. simple speech, environmental sounds, smell, body signs, e tc.) in order to help him orientate in activity, to understand what is going on, as well as to build his confidence and participation. From my privies observations of Pupil Z, I have also implemented motivating stimuli and activities during the two-week observation programme in order to motivate him (Appendix, 4). I have used the school timetabling curriculum forms for key stage 2 (see Timetable 2 and 3) to present my findings. The observation gathered on the observation form, (timetabling curriculum forms) were filled in traffic lights; green, orange, red and blue (Table 2 and 3). This overall description of the alertness categories of Pupil Z was complemented in agreement with class staff, alongside example behaviour for each category where Pupil Z displayed most common behaviour during school days. Table 1 Descriptions of Pupil Z different alertness levels Alertness level Colour Description Behaviour Active, focused on the environment Green Engaged in sensory activities (e.g., li stening, touching, smelling, etc.). Focused on other individuals or on materials in the classroom. Eyes open, head movements, body activity, reaching toward / for object(s), smiles, laugh and made some other types of vocalisation. Inactive, withdrawn Orange Focused on himself or without any focus. Activities are not directed to the environment. Flat posture, head down or turn

Discuss the analytical foundations of the Phillips curve Essay

Discuss the analytical foundations of the Phillips curve - Essay Example most of the policy makers believed that unemployment and inflation could not occur at the same time and whichever occurred, it became a major determinant of the policies that would be adopted by a given economy. Another issue that the economists believed was that it was possible to achieve one objective without having negative implications on the other. However, the research that Philips undertook in 1958 provided information that generated quite a number of questions to those assumptions. On his part, Philips used the available UK data to make an analysis on the rates of inflation and unemployment in UK. After plotting the result of the research, Philips noted that there existed an inverse relationship between wage inflation and unemployment. As a way of checking the relevance of the Philips conclusion, other economists used price inflation instead of wage inflation and noted that the results were similar curves and this became the origin of the Philip curve. This paper seeks to dis cuss the analytical foundations of the Phillips curve as well as how unemployment rate of 5% may be inflationary in one country, while it is deflationary in another country. According to the curve, any change in the level of unemployment produced a predictable as well as direct impact on the level of price inflation. Philips, together with other policy makers adopted the relationship between the two variables and noted that an increase in aggregate demand as well as fiscal stimulus in the economy triggers various responses. First, they noted that an increase in demand as the spending by the government rises results into national growth. This is followed by falling on the pool of unemployed. On their part, firms will be involved in competing for fewer workers available in the economy by increasing their nominal wages making the workers to have a higher bargaining power as they seek to have their nominal wages raised further (Forder, 2014). As the economy gets near to full capacity,

Organisations and Behaviour Essay Example | Topics and Well Written Essays - 2500 words - 2

Organisations and Behaviour - Essay Example In an organization where such culture exists emphasis is given on trust rather than orders (Grint 2005). This type of organizational culture has the following advantage: it can help to keep communication and cooperation with organization at high levels. Still, there is always the chance that certain employees take advantage of this culture for ignoring their obligations, according to their position; b) Role: the specific type of culture is based on bureaucracy; emphasis is given on rules and orders of superiors while hierarchy is highly respected. In organizations with such type of culture it is quite difficult for severe failures to exist at the level that employees’ activities are fully aligned with the rules set by the top management team (Grint 2005). On the other hand, this type of culture can harm team-work and communication within the organization at the level that employees would not feel as members of a team but as followers who have not the right to state their view in daily business operations; they have just the obligation to follow the orders given to them. In such environment employee performance cannot be easily kept at high levels; c) Task; it is the organizational culture that promotes the use of project-type tasks within each organization (Grint 2005); this type of culture promotes decentralization, leaving room for employee initiatives, a practice that can significantly enhance employee performance; however, this type of culture can set the integration of a business in risk if roles are highly decentralized; and d) Person: it is an organizational culture that highlights the value of person, as member of an organization (Grint 2005); such culture is applicable in family-owned firms or small firms but it cannot be used in large businesses (Grint 2005). 1.2 In the context of a business, the term structure is used in order to show ‘the form of an organization meaning especially the hierarchy of positions and relationships’ (Kn ights and Willmott 2007, p.197). Organizational structure, as described above, can be divided into three categories: a) Tall structure; it is met in organizations where the levels of hierarchy are many (Christensen et al. 2007); such structure can be useful for ensuring the control by the top management team of all business activities but it can threaten trust and communication; b) Flat structure; it refers to organizations with just one or two levels of hierarchy (Christensen et al. 2007); such structure enhances communication and team-work but it is difficult to be applied in large organizations; c) Matrix; the term denotes the dependency of each employee by many superiors, simultaneously; such structure is more appropriate for organizations in the public sector while in the private sector it could not be applied at the level that in the private sector independent departments usually exist for covering business needs (Christensen et al. 2007). On the other hand, in small businesse s such structure could be possibly applied. 1.3 The ‘chain of command’ principle is based on the idea that within organizations the full independency of individuals is not acceptable, meaning that ‘each employee should necessary have a superior to whom he had to report’ (O’Fallon and Rutherford 2010, p.76). This principle is applicable in organizations since it does not set as a prerequisite the excessive power of the superior over the employee but just the like the dependency of the employee

Designing marketing campaign for ExxonMobil Qatar Coursework

Designing marketing campaign for ExxonMobil Qatar - Coursework Example The main target groups of the marketing strategies are the customers, the shareholders, business partners and host governments. Some of the marketing strategies employed here include advertising that is done through print media, car stickers, TV commercials as well as internet based advertisements. The TV commercials serve to complement the print media such as magazines, newspapers, and business journals. The adverts have the company profile and the financial programming. They are run on the Arabic networks such as JSC and MBC.The Company has also been able to organize sports as a way to make it known for the products as well as the services it offers in the market. Some of the sports have included the organization of tennis, hockey and golf tournaments that are usually based in Doha ("ExxonMobil Qatar Inc," n.d.). The events are organized by the Qatar Olympic committee that oversees the participation. The company has also been supporting science and technology in the schools by recruiting some of the best University students in Qatar into the company making it well known all over the region. The company has also been able to embrace technology and has now been able to market its products through the internet marketing strategies through their websites as well as social networking sites such as twitter, Face book among others ("ExxonMobil Qatar Inc," n.d.). The company has also been engaging in activities that are in tandem with corporate social responsibility they have been able to promote safety awareness in the region through what they call the pink t-shirt campaign. Through this the company supports relief services all over the world. The business women an award that acknowledges successful women is also some form of marketing Campaign. Through the marketing campaigns the company has been able to reach its customers in the region. Requests for partnerships have also been on the rise, since the inception of various marketing strategies. Most

Constitutional Rights before and after Arrest Research Paper

Constitutional Rights before and after Arrest - Research Paper Example The sixth Amendment done to the Constitution of United States is the part of the Bill of Rights which put forward privileges associated to the prosecutions of the criminals. An arrest causes the withdrawal of an individual’s freedom by a police man or any individual with an officially authorized permission. Usually an arrest takes place when a person is alleged of carrying out a crime and therefore should be taken into custody. â€Å"Because an arrest is in effect a â€Å"seizure† it must conform to the probable cause and warrant requirements of the Fourth Amendment†(Otis.H.Stephens). According to the author arrests are frequently made in accordance to warrants build on initial inquiries so it is given out by a judge on apparent grounds. A court permits the investigation of a criminal activity by issuing a court order also known as writ. Therefore any investigation or arrest without a legal warrant is unfair and unconstitutional. A person has the right to demand as to why he is being arrested and the kind of allegation against him. An individual must see the warrant to be certain that his name exists on it and to confirm the charges. Moreover the person has the right to be informed of his constitutional rights also known as â€Å"Miranda rights† before he is interrogated after the arrest. These include the right to remain silent and refuse to cooperate without the consultation of your attorney as the police can utilize those answers against the person seized in the court. If an individual doesn’t meet the expense of a lawyer, he has the privilege to get a public defender without any cost and discuss his case with the attorney before further proceeding with the investigation. A person can also say no to every physical or medical test such as breathalyzer, lie detector etc. he also has the permission to contact any relative or important person to inform them about his arrest and so is allowed to make telephone calls. A person ar rested with no warrant is required to be carried quickly before a court official for a credible hearing of the case. Most of the times suspects defend against arrest therefore the police might use physical force or violence to take them in custody. â€Å"The courts have generally recognized that the fourth amendment permits police to use only such force as is â€Å"reasonable† and â€Å"necessary† in effectuating an arrest† (Otis H.Stephens). Therefore the person being suspected should give in without much resistance and has the right to take legal action against the police if they violate this amendment. The fourth amendment has forbidden unfair investigations and seizures of individuals and their possessions without a legal warrant specifying the place to be explored or the person to be arrested. Now as discussed earlier the Fifth Amendment permits an individual to refuse giving a testimony that might convict himself both in judiciary and in answer to interrog ation by the police. It also contains the clause of â€Å"Double Jeopardy’ in which it is not allowed to condemn a person twice for the same allegation. The sixth amendment however points out rights that are relevant in every criminal trial such as the right to an immediate and open hearing by an neutral panel