Locating The Xhoi Recognition Site On Lambda Dna Using

Locating the XhoI Recognition Site on Lambda DNA Using a HindIII and XhoI double digest Haleigh Wood Abstract Restriction enzymes cut DNA at certain sites to create multiple DNA fragments. Restriction enzyme HindIII has known DNA fragment lengths and recognition sites when digesting lambda DNA, while the lambda DNA recognition site for restriction enzyme XhoI is unknown. The goal of this study is to determine the lambda recognition site of XhoI by comparing a HindIII digest and a HindIII and XhoI double digest on an electrophoresis gel. The HindIII digest had a band at 9.4 kb, but this band was not visible in the double digest, therefore we concluded the recognition site for XhoI was around 9.4kb. There were also two additional DNA†¦show more content†¦XhoI is a type I restriction enzyme, so the location of the recognition site on lambda DNA is unknown but it is known that it cleaves at CTCGAG sites (XhoI (10u/ul)). While the DNA fragment lengths and cleavage sites are known for HindIII lambda DNA digest, they are not known for a XhoI lambda DNA digest. The goal of this study is to determine the recognition site of XhoI on lambda DNA by comparing the DNA fragments from a HindIII digest to the DNA fragments of a HindIII and XhoI digest. We will do this by creating a mixture of solutions containing only DNA, DNA and HindIII, DNA and XhoI, and DNA, HindIII and XhoI. We will then run a gel electrophoresis, which will separate the DNA, fragments by size and we will compare the DNA fragments from each solution. Through comparing the DNA fragments from the HindIII digest and the HindIII and XhoI double digest, we will be able to determine the XhoI recognition site on the lambda DNA. Methods Preparation of restriction digest solutions: To begin the process to determine the XhoI recognition site in the lamda DNA fragment we first prepared 4 tubes of solutions containing 10X Optizyme reaction buffer, sterile water, lambda DNA (0.3 ug/ul), XhoI (10u/ul, 3000u), and HindIII (10u/ul, 7500ul). Tube 1 contained 2ul 10X Optizyme, 16ul sterile water, and 2ul lambda DNA. Tube 2 contained 2ul 10X Optizyme, 14ul sterile water, 2ul lambda DNA, and 2ul XhoI. Tube 3 contained 2ul 10X Optizyme, 14ul sterile water, 2ul

Arguments For Favor Of Marriage - 969 Words

Her arguments in favor of marriage, though demonstrating a hearty common sense, are also suspect -- while it is true that marriage peoples the earth and replenishes existing stocks of virginitee, her own marriages do not seem to have produced any offspring, and while it may be her marriages, despite her claim that, do not seem to have prevented her from which is after all what marriage was, according to her, supposed to prevent. Moreover, from the account she gives of her marriages, it becomes increasingly obvious that marriage (with her, at least) is not quite so beneficial as one might think -- the only benefit the husbands get, in exchange for their purgatorie, is that of her with the possible exception of Jankin, who satisfied her better than but it is worth observing that she never speaks of the sexual act as giving the male partner pleasure on the contrary, she speaks of the husband s dette to his wife, Also, while she claims Biblical support for her views on marriage, the support that she cites is conveniently edited to suit her purposes (for example, Solomon did have 700 wives and 300 concubines -- but his appetites led to his turning away from God; and the marital relationship specified in the Bible is a reciprocal one rather than the one-sided one she speaks of, tilted in favour of the wife -- she conveniently ignores that while the he also exhorts women to love their husbands), and she elsewhere ignores the Bible when it proves difficult to gloseShow MoreRelatedThe Case : Great Promise And Its Impact On The Future1269 Words   |  6 Pagesthe subject of marriage equality. 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Colorectal Cancer for Inflammatory Bowel Disease - myassignmenthelp

Question: Discuss about theColorectal Cancer for Inflammatory Bowel Disease. Answer: Colon and rectum tumors are mostly common; the colorectal area is now the third most common site of cancer cases and deaths globally (Bailey and You, 2014). Cancer of the colon and rectum develops from the epithelial lining of the intestine. It usually begins as a benign polyp which later becomes a malignant which penetrates and destroys the normal tissues. The frequency of occurrence is highest for individuals older than 85 years of age and is higher for individuals who have had a history of colon cancer in the family. The specific cause of colorectal cancer is still not known but the risk factors have been identified. The risk factors include breast cancer in women, high fat and beef intake, and medical record of inflammatory bowel disease and lastly family history of colon cancer (Ponteri-lewis, 2013). If this disease is detected early and treated, it could save three people out of four with colon cancer. This essay is going to talk about the drugs administered to Nelly their mech anism of action and lastly the technologies used to detect colorectal cancer. Drugs administered Colorectal cancer therapy depends on the stage of the disease and it usually done by removal of the tumor through surgery, adjuvant therapy and supportive therapy. Nellie had a stage IVA cancer and she was referred to a colonoscopy and it was discovered that nelly had polyps in the colon which were surgically removed. She later came for adjuvant therapy which included 5-Fluorouracil, Irinotecan and lastly Oxaliplatin. This kind of therapy was used to reduce the chances of the cancer coming back after surgery. Pain medication was initiated after targeted radiotherapy failed to shrink the tumor, the severe back pain was as a result of cancer that had spread to the spinal cord and was compressing the spinal cord. Pain medication included paracetamol and morphine. Spinal tumor increased each day which caused more pain and the dose of morphine was difficult to titrate due to severe side effects it caused. End of life pathway drugs were commenced which included levomepromazine and sublingu al clonazepam. Mechanism of action 5- Fluorouracil (5-FU) is an antimetabolite which exerts its action on cells undergoing cycling. It kills tumor cells which are in the cycling and resting phase of the cell cycle. In the cell fluorouracil is changed to 5- fluoro-2-deoxyuridine-5-monophosphate (5-FdUMP), which causes the suppression of thymidylate synthase which later leads to the death of thymineless of cells. DNA synthesis is inhibited by the penetration of FdUMP. RNA synthesis and function is inhibited by 5-fluorouradine-5-triphosphate (FUTP) which is a fluorouracil metabolite. In the tumor cell the mechanism of resistance include an increased thymidylate synthase activity, a reduced activation of 5-FU, and lastly a decreased drug sensitivity of this enzyme. As a result the cell is unable to divide and dies. Irinotecan is a plant derived cell cycle specific (CCS) drugs which is found in the camptothecins group. It is converted to an active form SN-38 which requires carboxyl esterase (Huangg, Wuerzberger-Davis and Seufzer, 2012). it inhibits topoisomerase 1 activity hence causing damage to the DNA. The DNA is damaged by an inhibiting enzyme that cuts and the single DNA strands during the normal DNA repair process (Maureen, Beidler and cheng, 2013). Oxaliplatin is a chemotherapeutic agent that contains 1, 2- diaminocyclohexane ligand. Like the other platinum based compounds it exerts its effect by damaging the DNA and also by inhibition of RNA synthesis. It induces three types of links which include DNA intra-strand link, DNA inter-strand crosslinks and DNA-protein crosslinks. Intra-strand links act by the induction of DNA lesions while the inter-strand link are said to bring about the cytotoxicity of cisplatin. The DNA protein crosslink causes arrest of DNA synthesis and inhibition of messenger RNA synthesis hence cell death. Paracetamol also known as acetaminophen is a weak inhibitor of cyclooxygenase-1 (COX-1) and COX-2 are inhibitors of prostaglandin (Boataud, Aronoff and Richardson, 2015). Prostaglandin is inhibited if there is a low level of arachidonic acid. Cox-2 is the main isoenzyme when the arachidonic acid is at low concentration (Aronoff, Boutard and Marnett, 2013). And for this reason paracetamol selectively prevents prostaglandin synthesis involving Cox-2 this is due to the lower flux in the pathway that produces decreased level of hydroperoxide. Opioids acts on the receptors found in neuronal cell membrane to produce effect. The major three types of receptors include kappa, delta, and mu. Morphine has been found to have a high affinity for m receptors. G-protein which is a guanine nucleotide protein is coupled to opioid receptors. The opioid has two sites where it acts upon, the postsynaptic neuron and the presynaptic nerve terminal. In the presynaptic nerve terminal the action of morphine is to prevent neurotransmitter release by reducing calcium entry and increasing potassium current. Morphine also acts on the pain pathways, opioid receptors are present in pain transmission regions of the nervous system. It acts by inhibition of neurotransmitter release from the primary afferent terminals found in the spinal cord and also the activation of the midbrain control that is the descending inhibitors. Levomepromazine is a neuroleptic agent. It has anti-histamine, anti-emetic and analgesic effect. It precise mechanism of action is not yet known. Clonazepam is a benzodiazepam. It is a strong anticonvulsant. Its main type of action is to make it easier for GABAergic transmission in the brain. This GABA receptors are located in dorsal raphe neurons which acts by inhibiting raphe cell lining. 5-HT synthesis is not inhibited by clonazepam but it reduces the utilization of 5-HT in the brain. Technologies used to detect colorectal cancer Colonoscopy is a procedure in which one views the large intestine using a device known as a colonoscope (Zapka, Klabunde and Typlin, 2014). It is a thin flexible tube with a camera and a source of light. This device is inserted in the anus region and through the colon. This device captures images and videos for the doctor to see. If there is an abnormal growth the doctor will be able to see. It is also used to remove little amount of tissue for biopsy. DNA testing has been used to identify individual at risk of colorectal cancer. Through DNA testing doctors have been able to prevent the disease at an early stage. Clients identified at risk are genetically cancelled on the disease, so as to understand prevention and treatment measures (Leissic, 2012). Stool test has been used to check for the presence of blood. It is the most common method used for early cancer detection (Burch, Soares-Weiser and John, 2013). The basic examination of the stool includes checking for color and testing for occult blood. It also checks for the ion containing portion of the hemoglobin known as the heme which is changed during the movement in the intestines (Ouyang, Chen and Getzenberg, 2018). Lower gastrointestinal tract procedure has been used to detect the presence polyps and tumors in the colon. Barium enema is inserted in the rectum so as to visualize the presence polyps. This radioscope substance is inserted during fluoroscopy. Other visualizing colon studies include double contrast studies. In double contrast, it involves the insertion of thick barium solution followed by pumping of air. This process distinguishes barium coated mucosa and air filled lumen hence allowing easier detection of little lesions. Sigmoidoscopy, anoscopy and protoscopy are used to view the lower portion of the colon to check for rectal bleeding, tumor, polyps or other pathological processes. The anoscope is used to view the anus and the lower rectum. The sigmoidscopes and protoscopes are rigid scopes used to view the rectum and sigmoid (Elmunzer, Harvard and Schoenfeld, 2015). The flexible fiberoptic sigmoidoscope allows the colon to be viewed up to 40-50cm from the anus. During the procedure biopsy can be performed using a small biting forceps introduced through the endoscope. If the rectal polyps are viewed that can be removed with a wire snare used to grasp the pedicle. Magnetic resonance imaging is used in the checking of polyps in the colon (Forsberg, 2012). This is a non-invasive technique that uses radio waves and magnetic fields to produce an image of the area being studied. Oral contrasts agents are used to enhance images. The client lie in the machine having not eaten or drunk anything for the last 6-8 hours. The entire procedure takes up to thirty to ninety minutes. Scintigraphy is a radionuclide testing which uses the radioactive isotopes such as the indium and iodine. This agents are used to reveal or visualize changes in atomic structures, organ size and the presence of neoplasm or any other focal lesion. Abdominal ultrasonography is another method used to check polyps in the colon. This is done by production of high frequency sound waves in the internal body organs. This echoes are recorded on an olliscope as they pass through the tissues. This method is used to check the size of and the configuration of abdominal structures. Advantages of this method is that it needs no ionizing radiation hence they will be no side effects and it also not expensive. The only disadvantage that it has is that it cannot view structures behind a bone since the bone prevents ultrasonic echoes from passing to the deeper structures. References Aronoff, D., Boutard, O. and Marnett, J. (2013). Inhibition of prostaglandin H2 synthases by salicylate is dependent on the oxidative state of the enzymes.J. Pharmacol. Exp. Ther, 304, pp.589-594. Bailey, E. and You, N. (2014). Increasing disparities in the age related incidence of colon and rectal cancer in the United States. 150(1), pp.17-22. Boataud, O., Aronoff, D. and Richardson, J. (2015). Determinants of the cellular specificity of acetaminophen as an inhibitor of prostaglandin H2 synthases.Proc. Natl. Acad. Sci. USA, 99, pp.730-735. Burch, T., Soares-Weiser, K. and John, D. (2013). Diagnostic accuracy of fecal result blood tested in screening colorectal cancer: A systemic review.journal of medicine screening, 14(3), pp.132-137. Elmunzer, B., Harvard, R. and Schoenfeld, P. (2015). Effect of flexible sigmoidoscopy based screening on incidence and mortality of colorectal cancer: A systematic review and meta-analysis of randomized controlled trial.Plus Medicine, 9(12), pp.1001-1035. Forsberg, F. (2012). Advance in ultrasound contrast technique.Applied referencing radiology, 16, pp.5-10. Huangg, T., Wuerzberger-Davis, S. and Seufzer, B. (2012). NF-Kaapab activation by capmtothelin, a linkage between nuclear DNA damage and cytoplasmic signed events.J clin invest, 107, pp.241-246. Leissic, M. (2012). Advance in genetic testing for care risk.Medsurge nursing, 10(3), pp.123-125. Maurren, J., Beidler, R. and cheng, Y. (2013). Campthocesin resistance related to to drug induced down regulation of topoisomerase 1 and steps occurring after the formation of protein linked DNA breaks.AnnNy Acad Sci, 803, pp.74-92. Ouyang, D., Chen, J. and Getzenberg, R. (2018). Non-invasive testing for colorectal cancer: A review.American journal of gastroentology, 100(6), pp.1393-1403. Ponteri-lewis, V. (2013). Colorectal cancer.Medsurge nursing, 9(1), pp.9-15. Zapka, J., Klabunde, C. and Typlin, S. (2014). Screening colonoscopy in the USA: Attitudes and practises of primary care physians.Journal of general intervid medicine, 27(9), pp.1150-1158.

Thomas Jefferson Essay Example

Thomas Jefferson Essay Thomas Jefferson and His Effect on America Thomas Jefferson possessed one of the greatest leading minds of colonial America. Literate in political theory, scientific farming, natural history, and architecture, Thomas Jefferson personified the optimistic spirit of Enlightenment thinking. Thomas Jefferson proved to be one of the major forces in the founding and developing of America. When Jefferson became the third President of the United States, he immediately made an impact on the lives of average Americans by reiterating his idea of a Jeffersonian Republican Yeoman Farmer. Leading the nation, Jefferson made significant changes in the government, spurred the idea of American movement westward, and worked to fix the ever-growing slavery issues and better the education system to spring America forward and develop it into the prosperous nation it is today. Prior to Jeffersons presidency, Jefferson succeeded Benjamin Franklin as the minister to France in 1785. As Secretary of State, Jefferson took part in both domestic and world affairs. Jefferson clashed frequently on many issues with Secretary of the Treasury, Alexander Hamilton (Bernstein 101). The conflict between Jefferson and Hamilton caused many other disagreements in American politics that pinned former allies against one another. With the government on opposite ends, two political parties began to form: the Federalist Party lead by Hamilton and the Jeffersonian Republicans lead by Jefferson. The Jeffersonian Republicans were opposed toa strong central government whereas the Federalists favored the strong central government. Jefferson preferred a less centralized government with more power for the states. We will write a custom essay sample on Thomas Jefferson specifically for you for only $16.38 $13.9/page Order now We will write a custom essay sample on Thomas Jefferson specifically for you FOR ONLY $16.38 $13.9/page Hire Writer We will write a custom essay sample on Thomas Jefferson specifically for you FOR ONLY $16.38 $13.9/page Hire Writer Jefferson, being a brilliant man, drafted the Northwest Ordinance also known as the Ordinance of 1787. This ordinance provided a structure for government of the Northwest Territory. Jeffersons outstanding leadership led to his being elected President of the United States in the 1800 election. Jeffersons most lasting contribution to America is the Declaration of Independence. Specifically chosen, Jefferson needed to carefully choose his wording so that the document would be persuasive enough towards a number of parties (The Declaration of Independence). Congress intended for Americans to read it and want to become a part of the patriot cause and foreign powers read it and want to aid the ilitia. In the first part of the Declaration of Independence, Jefferson expresses key concepts at the core of American beliefs such as all men are created equal, unalienable rights, and life, liberty, and the pursuit of happiness. In the next section, Jefferson listed injustices that the colonies considered independence worthy. Jefferson accused King George of trying to construct absolute tyranny in America so in the conclusion, Jefferson officially destroys all ties with Britain and is now guilty of treason. Everyone in the congregation could be condemned to the gallows if rosecuted before a royal court. By a vote of twelve to zero, the colonies approved the document and declared independence from Britain on July 4, 1776. One of Jeffersons greatest achievements as President includes his investment on the Louisiana Territory owned by the French. Jefferson visualized American westward expanslon ana took advantage 0T tne opportunlty presented to nlm. Napoleon Bonaparte, Frances ruler, needed money to fund the war against the British so he offered all of Frances claims in North America to Jefferson. With the territory tretching a vast 820,000 square miles, the transaction would amount to about three cents per acre totaling a whopping $15 million dollars. However, Congress originally only approved to spend $10 million, so the purchase sent the nation into a large debt. This famous transaction came to be known as the Louisiana Purchase. The United States, now doubled in size, grew beyond the Mississippi River to include forests, plains, prairies, and mountains. Surprisingly, Jefferson did not intend to buy all of the Louisiana Territory, Just the port of New Orleans so that the United States, ot its competitors such as Spain, France, and Great Britain, could control the mouth of the Mississippi River (De Cesar). With the Louisiana Purchase fulfilling his dreams of westward expansion, Jefferson now sought to launch an expedition to explore the new addition to America. Minus what French traders and fur trappers and Spanish and British explorers shared about the west, nobody really knew what the western part of the country held. Jefferson assigned a prominent frontiersman named Meriwether Lewis to lead the U. S. Army expedition (known today as the Corps of Discovery). Then Lewis hired William Clark to accompany him on the Journey, hence the name The Lewis and Clark Expedition. Jefferson sent a letter to Congress asking for $2,500 to fund the trip through the Louisiana Territory and on to the Pacific Ocean. Jeffersons main goals included establishing trade with the Native American people of the West as well as to find a straight shot water route to the Pacific (Thomas Jefferson and the Lewis and Clark Expedition). Jefferson also assigned Lewis and Clark the task of documenting the geography of the West, how the Native Americans communicated and lived, and ow things like the plants and animals, the soil, the rocks, and the weather differed from those in the East. The total round trip lasted two years and four months and the work of Lewis and Clark and the Corps of Discovery instituted an American presence all across the continent. A major problem Jefferson faced during his Presidency involved the Barbary Pirates from the Barbary States of Tripoli, Tunisia, Algeria, and independent Morocco. These pirates harassed and preyed on the commercial shippers that traveled through the Mediterranean Sea. Soon debate began on how to deal with these irates. After an American brig faced capture by a Moroccan pirate ship in the Atlantic Jefferson stressed, Our trade to Portugal, Spain, and the Mediterranean is annihilated unless we do something decisive. Tribute or war is the usual alternative of these of pirates Why not begin a naw then and decide on war? (Roberts). In 1801 Jefferson refused to pay tribute to the pirates and deployed a U. S. Navvy force to the Mediterranean. If the United States wished to continue their commercial trade, they would need a protective force at sea (Roberts). The U. S. Navvy took military ctions against pirates by blockading the Barbary ports to achieve a deadlock and included an American land assault to gain control of the main city. By 1805, Jefferson signed a treaty ending the Barbary Wars and made peace with the Barbary States. LIKe most wealtny American men In Jeffersons time, ne owned slaves. However, Jefferson opposed slavery his whole life and felt it went against the laws of nature. He knew everyone had a right to personal liberty (Thomas Jefferson and Slavery). Jefferson worked to abolish slavery but did not have strong political support. When rafting and proposing many ordinances, he tried somehow to work in the ordinance banning slavery. When he drafted the Northwest Ordinance, it outlawed slavery in the Northwest Territories. Jefferson continued to support abolition but slavery continued to grow because the demand for laborers increased. To try and end Virginias support towards slavery, Jefferson encouraged growing crops that required little slave labor. If the abolition of slavery did not happen, Jefferson feared the federal union would be destroyed by slavery and result in a civil war that could tear he nation apart (Thomas Jefferson and Slavery). Jefferson became deeply concerned the next generation of leaders would not end slavery. One of Jeffersons lasting legacies and greatest accomplishments is the founding of the University of Virginia. Ideas of educating the common man occupied Jeffersons mind. In a letter to James Madison, Jefferson wrote, Above all things I hope the education of the common people will be attended to; convinced that on their good sense we may rely with the most security for the preservation of a due degree of liberty. (Quotations on Education). Jefferson developed much of the University and designed the curriculum, secured its funding, and generated how to hire teachers. The University of Virginia is a public university funded by the state of Virginia. Jefferson intended the university to be dedicated to teaching and grooming future leaders. Jefferson believed they needed to be educated in practical affairs and public services (Founding of the University). Jefferson retired from office in 1808 but still continued to contribute to the nation. When speaking with James Madison, Jefferson said he wanted to be remembered as wo things: the author of the Declaration of Independence and the founder of the University of Virginia (Kindig). Jeffersons wishes came true because this is what Jefferson is most famously known for. Overall, Jefferson created a positive lasting effect on America. Today in Washington D. C. there stands an enormous 19-foot statue of Jefferson that is surrounded by passages from the Declaration of Independence as well as passages from his other writings. The Jefferson Memorial stands as a timeless monument dedicated to one of Americas most influential Founding Fathers. Thomas Jefferson Essay Example Thomas Jefferson Essay Thomas Jefferson faced a moral dilemma over the purchase of the Louisiana Territory from France. There were as many advantagesto the Louisiana Purchase for President Thomas Jefferson as there were disadvantages. In reference President Thomas Jefferson believed that the United States should be a nation of independent farmers. When France offered to sell the Louisiana Territory to the United States in 1803, Thomas Jefferson wanted to seize the opportunity to double the size of the nation and to provide future generations with a supposedlynever-endingsupply of new farmland. ButThomas Jefferson was a strict constructionist; He believed that the federal government had no powers other than those specifically listed in the Constitution; and the Constitution did not authorize the president to buy territory from foreign nations. (https://www.gilderlehrman.org/history-by-era/age-jefferson-and-madison/resources/louisiana-purchase-0) The main disadvantages and moral dilemma that Thomas Jefferson faced resided heavily in political reality. Thomas Jefferson had always advocateda very strict platform of Republican values.. This position had been seen early on in his disagreements with Alexander Hamilton in PresidentGeorge Washingtons cabinet.In the election of 1800, Thomas Jefferson was able to express a new type of government that influenced Republicanism.Thomas Jefferson took office and carried himself as the embodiment of Republican values.A significant aspect of this viewed limited government as essential.Republicans believed that the role of government needed to be decreased.Thomas Jefferson sought to minimize federal control through reducing its span.He pursued a laissez- faire approach to governance in his time as President and embraced it as the natural leader of the Republican party. We will write a custom essay sample on Thomas Jefferson specifically for you for only $16.38 $13.9/page Order now We will write a custom essay sample on Thomas Jefferson specifically for you FOR ONLY $16.38 $13.9/page Hire Writer We will write a custom essay sample on Thomas Jefferson specifically for you FOR ONLY $16.38 $13.9/page Hire Writer The Louisiana Purchase strained this belief in limited government.In the act of the purchase, Thomas Jefferson was increasing the po