Isolation and Characterization of Onion DNA

Isolation and Characterization of Onion DNA The experiment was about the isolation and characterization of DNA. The DNA was isolated from the onion. The mass of the isolated DNA was 15.11 g. The purity of isolated DNA was estimated by calculating the ratio based from the absorbance at 260nm and 280nm resulted to 0.671 meaning more protein was absorbed. Meanwhile in denaturation of DNA, the initial absorbance at 260 nm was 1.304 higher than the absorbance at 260 nm after heating which was 1.095. INTRODUCTION Deoxyribonucleic acid (DNA) is the genetic material in humans and all other organisms. DNA isolation is the removal of DNA from the cell which it normally resides. Isolation is the removal of DNA from the cell in which it normally inhabits. (1) Onions are used since it contains little amount of starch which allows the DNA to be more visible. The filtrate is made up of onions treated with salt, distilled water and detergent collectively called as lysis solution. DNA purification is done by enzymatic degradation of contaminating proteins with ethanol. A spectrophotometer is used in determining the concentration and purity of the proteins. (2) MATERIALS AND METHODS Isolation of DNA from Onion The peeled onion bulb was chopped and measured homogenized. The sample was placed in a blender added with an ice-cold lysis solution then for 45 seconds at low speed. Meanwhile, the lysis solution used was prepared beforehand by mixing 5.00 ml of liquid detergent, 5.00 ml of 0.500M EDTA, 10.0 ml of 50% Na Cl solution, and 80 ml of distilled water and placed in an ice bath. After homogenizing, the sample was filtered through the cheesecloth and the collected filtrate was placed in a 250-ml beaker. A 10.0 ml of 5% pepsin solution was added to the filtrate and placed on an ice bath for 10 minutes with occasional stirring. Ice cold 30.0 ml of 95% ethanol was pipette to the side of the beaker containing the sample and stand for 10 minutes on ice bath. Once the DNA precipitates appeared at the interface of the solution, the DNA was already ready for isolation. The spooled DNA was transferred immediately to a pre-weighed 100-ml beaker to determine the mass and percent yield of the sample. T he isolated DNA was added with 10.0 ml of 95% ethanol then covered with aluminum foil and refrigerated in preparation for the next laboratory procedure. Characterization of DNA Little amount of DNA sample was placed in a test tube added with 1.00 ml of 20% TCA followed by heating the sample for 10 minutes in water bath with 1.00 ml distilled water. A 2.00 ml of diphenylamine solution was added then heat again in a water bath for 10 minutes. The color change was observed and the absorbance of the sample from 400 nm to 700 nm was scanned to determine the wavelength of maximum absorption. Mean while, little amount of the DNA sample was placed in a separate test tube filled with 5.00 ml distilled water and scanned to read the absorbance at 260 nm then at 280 nm. After determining the A260/A280 value, the sample was heated to boil for 5 minutes and read the absorbance adain at 260 nm. RESULTS AND DISCUSSIONS The mass of the raw sample gathered from onion is 30.4 g. After homogenization and adding of pepsin solution and ethanol, DNA precipitates were became visible and transferred to another beaker. The isolated DNA measures 23 g. The calculated percentage yield was quite high. However, still some sources of error was done while conducting the experiment, the sample with DNA precipitates was disturbed while transferring the DNA. The accumulated DNA precipitates is enough for the next procedure which is characterization. Heat denaturation of DNA, causes the double helix structure to unwind and form single stranded DNA. Thus, the bases unstacked and can absorb more light causing an increase after denaturation. But based on the results gathered, the initial absorbance at 260 nm was 1.304 then was decreased after heating which was 1.095. The calculated percent increase in absorbance was 8%. This error is maybe, due to the heating process. The DNA acquired was quite greater and was not totally heated afterwards causing double helix structure not to unwind and form a single stranded DNA. The filtrate gathered from this experiment was made of onions and lysis solution. Onion was used in this study due to low starch content, allowing the DNA to be more visible considering the onion as one of the best source of DNA. (4) The used of lysis solution was to separate the DNA from extra cell components and to keep the location in which the DNA will not be tainted. The NaCL provides NA+ ions that will obstruct the negative charge as of phosphate ends of DNA. Permitting these ends to come nearer so they can precipitate out of a cold solution. The detergent causes the breaking down of the cell membrane by emulsifying the cell proteins and lipids. Also, disrupting the polar connections that collectively holds the cell membrane. The complexes formed with these lipids and proteins causes the precipitate out of solution. Meanwhile, the purpose of EDTA is to chelates metal ions. (5) A Pepsin solution was used for purification via enzymatic degradation. DNA is polar due to its extremely charged phosphate backbone which makes it soluble in water. Thus DNA is insoluble in ice cold ethanol, as a result when the cold ethanol was added, it causes stable ionic bonds to form and precipitate the DNA. Heating the sample is the one responsible for the formation of the observed color of DNA with diphenylamine. When the DNA is heated with acid, the 2-deoxyribose is converted to w-hydroxylaevulinic aldehyde, which reacts with the compound diphenylamine. Through this, a blue-colored compound supposed to produce. In our sample the color observed was green possibly because of the DNA concentration. The ratio of absorptions at 260 nm vs 280 nm is frequently used to evaluate DNA contamination of protein solutions. The nucleic acids, DNA and RNA, absorbs at 260 nm and proteins absorb at 280 nm. Based on the results, the rate ratio of absorptions at 260 nm vs 280 nm is 0.671. Since proteins absorb light at 280 nm, the ratio is low meaning there is a lot of protein absorbed at 280nm.

Edgar Allan Poe :: Essays Papers

Edgar Allan Poe2 Edgar Allan Poe’s life had a profound effect on the technical style of his writing. Poe spent most of his life raised by foster parents who did not contribute to or encourage his writing. His first work was published in 1827, at the young age of 18, but his story in 1833, â€Å"MS Found in a Bottle,† marked the beginning of his writing career. Poe uses darkness and death in many of his stories. In his poems he was able to illustrate moods of mystery very well. Edgar Allan Poe was a unique writer who was not afraid to do something different from all the authors, while his adulthood was short and depressing. Born to traveling actors David and Elizabeth Poe on January 19, 1809, Edgar Poe was the middle child of three children. His father, David Poe, was from a Baltimore family. He was an actor by profession and a heavy drinker. Edgar was never very close with his older brother, William Henry Leonard Poe, because he had been left with his paternal grandparents around September 1807 for what began as an undetermined amount of time. In July of 1810 David Poe deserted his family and died shortly after. His death was most possibly alcohol elated. Elizabeth was still pregnant with their youngest child, Rosalie, who was born that December, at this time. Approximately a year after her daughter was born, in December of 1811, Elizabeth Poe died of tuberculosis. As a result of her death, William Henry Leonard stayed with his grandparents, Edgar was adopted by a couple of wealthy merchants from Richmond, Virginia, John and Fanny Allan, who offered him a better education than his grandparents could , while Rosalie was adopted Mr. and Mrs. William Mackenzie. In 1815, John Allan moved the family to England to try to make Allan and Ellis prosper. While there, Edgar went to private schools where his creative writings were discouraged. In 1820 the Allan’s returned to Virginia as a result of the collapse of John’s business venture. Shortly after the Allan’s return to the United States, Edgar began to support himself as he moved to Boston and worked in a merchandise house. Edgar Allan Poe :: Essays Papers Edgar Allan Poe2 Edgar Allan Poe’s life had a profound effect on the technical style of his writing. Poe spent most of his life raised by foster parents who did not contribute to or encourage his writing. His first work was published in 1827, at the young age of 18, but his story in 1833, â€Å"MS Found in a Bottle,† marked the beginning of his writing career. Poe uses darkness and death in many of his stories. In his poems he was able to illustrate moods of mystery very well. Edgar Allan Poe was a unique writer who was not afraid to do something different from all the authors, while his adulthood was short and depressing. Born to traveling actors David and Elizabeth Poe on January 19, 1809, Edgar Poe was the middle child of three children. His father, David Poe, was from a Baltimore family. He was an actor by profession and a heavy drinker. Edgar was never very close with his older brother, William Henry Leonard Poe, because he had been left with his paternal grandparents around September 1807 for what began as an undetermined amount of time. In July of 1810 David Poe deserted his family and died shortly after. His death was most possibly alcohol elated. Elizabeth was still pregnant with their youngest child, Rosalie, who was born that December, at this time. Approximately a year after her daughter was born, in December of 1811, Elizabeth Poe died of tuberculosis. As a result of her death, William Henry Leonard stayed with his grandparents, Edgar was adopted by a couple of wealthy merchants from Richmond, Virginia, John and Fanny Allan, who offered him a better education than his grandparents could , while Rosalie was adopted Mr. and Mrs. William Mackenzie. In 1815, John Allan moved the family to England to try to make Allan and Ellis prosper. While there, Edgar went to private schools where his creative writings were discouraged. In 1820 the Allan’s returned to Virginia as a result of the collapse of John’s business venture. Shortly after the Allan’s return to the United States, Edgar began to support himself as he moved to Boston and worked in a merchandise house.

A Model of Christian Charity Essay

In 1630, John Winthrop led a fleet of 17 ships and a thousand puritans from England to the Massachusetts Bay. With a new place to call home, John Winthrop and the puritans hoped for a fresh start in the new world. The main reason for this new beginning was due to the fact that they had broken apart from England’s Church and had declared to start their own church. They believed that starting a new church was ordained by God and that the Massachusetts Bay area was given to them by God in order to start the Church. With a plan to start a new Church, John Winthrop created, â€Å"A Model of Christian Charity† to help set guidelines. These guidelines were a mix of logic and Biblical teachings that would be used to help start the new Church. Without these guidelines the new Church surely would have not been very successful. Winthrop was essential to the foundation of the new Church, and without him, the Church would have surely failed. John Winthrop’s writing gives a foundation in which the puritans can build the Church. A main point that John Winthrop wrote is that Christians need to start serving the Lord with all of their heart and try to increase the body of Christ. Increasing the body of Christ meaning that the puritans would influence others to trust in Christ and grow the population of Christ followers. The body of Christ means the community of those who follow Christ, and the doctrine that Winthrop wrote told the puritans that they need to profess themselves as followers of Christ. They need to be able to show others the Christ in them. Winthrop says that Christians are bound together by the love of Christ, and that love is unbreakable. Another main point that Winthrop mentions is that they need to go against the crowd, the crowd meaning the English Church. Since the puritans are breaking away from the English Church, then it would be wrong to copy the English Church. That would be too easy, instead they need to do what they feel is right and go against the English Church entirely. These main points of Winthrop’s covenant were vital to the foundation of the new Church. At the end of his covenant, Winthrop discusses what will happen if the puritans were to break the covenant. If the puritans were to break the  covenant then the Lord will strike them down for not keeping the covenant. Winthrop explains that Massachusetts would be a city on a hill in which people would admire and look up to. Not one that breaks the covenant and disobeys God, rather one that praises him. The puritans need to be a beacon of light to the world and show others the way to Christ. After the puritans had landed in Massachusetts, they began the new Church and succeeded in doing so. Later in history, people use Winthrop’s ideas to say that God had chosen the United States as a beacon of democracy and freedom for others to follow, just like people followed the puritans.

Anti Semitism And The Holocaust - 1682 Words

Imagine yourself as a seven year old child, playing hide and seek in the neighborhood streets with your mischievous friends and enjoying life without a care in your mind. Where you go to school with a smile on your face excited about what the day has to bring, messing around and cracking jokes with your friends at lunch time, and later on getting picked up by your nurturing parents to go home and do homework and have some family time with your parents and siblings and finally going to bed in your warm and comfortable bed. Now imagine if all those wonderful things just disappeared and the world around you changes in one instant and turns into a fiery pit of hell, where evil men roam those streets you found refuge. All you hear is an unending sound of screaming, crying, and gunshots as mothers, fathers, brothers and sisters are forever separated not knowing where they are going. Anti-Semitism is the hatred of Jews, and it reached its pinnacle during the Holocaust. It’s known wor ld wide as the genocide of approximately six million European Jews. Strikingly, there is a decent number of people in the world who don t entirely believe that the Holocaust happened as it is documented by witnesses, survivors, and those tasked with rescuing those who survived, and burying those who did not. These people are known as Holocaust Revisionists; or in another term, â€Å"deniers.† Farzan 2 Holocaust revisionists do believe that the Holocaust happened; however, they do notShow MoreRelatedAnti Semitism And The Holocaust876 Words   |  4 Pagesin the dark about it as well who could possibly blame America who was thousands of miles away to know. During the years of the holocaust a feeling of Anti-Semitism was clearly present all over the world this including the United States, this was seen in the results from different polls conducted and from the social policies at the time. These feelings of Anti-Semitism were convenient because it gave people an excuse to not do anything to help the Jews. Between 1938-1939 about 85,000 Jewish refugeesRead MoreAnti Semitism And The Holocaust1950 Words   |  8 Pages7th Period Holocaust Research Paper Anti-semitism is a concept that has been around in our society for a few centuries now, and has played a significant role in the world for many of those years. Although we see anti-semitism fade and diminish, its historical symbolism will hover over society for years to come. The Holocaust, taken place in Eastern Europe, is known to be one of the largest genocides this world has ever seen, and is the gruesome platform that people relate to anti-semitism. The HolocaustRead MoreWas German ‘Eliminationist Anti-Semitism Responsible for the Holocaust?832 Words   |  3 Pagesâ€Å"Was German ‘Eliminationist Anti-Semitism† Responsible for the Holocaust?† is a fascinating and somewhat discouraging debate that explores the question of whether German anti-Semitism, instilled within citizens outside of the Nazi Party, played a vast role in the extermination of Jews during the Holocaust . Daniel Jonah Goldhagen, author of â€Å"The Paradigm Challenged,† believe s that it did; and argues quite convincingly that ordinary German citizens were duplicitous either by their actions or inactionsRead MoreWas German â€Å"Eliminationist Anti-Semitism† Responsible for the Holocaust?2341 Words   |  10 Pages | |Was German â€Å"Eliminationist Anti-Semitism† Responsible for the Holocaust? | |Issue 10 â€Å"Taking Sides: Clashing Views in World History† | | | German anti-Semitism played the main role in Holocaust and extermination of Jewish population in Europe duringRead MoreAnti-Semitism and Lack of Concern Among Non-jews During the Holocaust997 Words   |  4 PagesAntisemitism is to blame for the lack of concern among non-Jews during the up rise of the Holocaust. nbsp;nbsp;nbsp;nbsp;nbsp;It is hard to grasp the number of lives lost during the Holocaust. How someone could have so much hatred towards one group of people. Or how so many people could set back and watch something like this take place without protest. To begin to understand how a tragedy like the Holocaust could have took place without intervention we need to understand antisemitism. nbsp;nbsp;nbsp;nbsp;nbsp;Merriam-WebsterRead MoreThe Tragedy Of The Holocaust Essay1118 Words   |  5 PagesWithin the era of 1933 to 1945, races struggled through times of gloom, torment and hopelessness. The Holocaust was hard on numerous people, with little survivors and still influences individuals present lives today. The majority of the anguish was brought about as a result of one man’s conviction, that had the capacity to impact a whole nation. The holocaust was a genocide that was exclusively centered around hatred found in Germany. Propaganda was one of the main causes, which conditioned the peopleRead MoreAdolf Hitler And The Holocaust Essay1105 Words   |  5 PagesAdolf Hitler and the Holocaust. 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It was an entirely fictional piece of work, but people believed the bookRead More Anti-Semitism Essay1369 Words   |  6 PagesAnti-Semitism Discrimination and prejudice have been in our world for as long as humans have themselves. Discrimination has caused problems in societies all throughout history. But despite all of the terrible things that have happened because of prejudice and discrimination, it continues to live on in our world today. Anti-Semitism, prejudice against Jews, is a form of discrimination that has caused perhaps the most problems throughout history. Many people describe anti-Semitism as more thanRead More The Origins of the Holocaust Essay1547 Words   |  7 Pagesorigins of the Holocaust has been studied by scholars using several differing approaches. These interpretations are outlined by Donald Niewyk in The Holocaust as the long history of European anti-Semitism, the charismatic personality of Adolf Hitler and the influence of modern â€Å"scientific† racism or eugenics. These interpretations are illustrated in the works of John Weiss, Ian Kershaw, and Henry Friedlander. Niewyk uses Weiss to identify the interpretation of ancient anti-Semitism located throughout