DNA Extraction from Strawberries; Alcohols

AbstractThis experiment investigated the gist of desoxyribonucleic acid extracted from strawberries. This was d unrivaled by implement the independent vari fit of intoxi s assoilt to affect the dependant covariant of the arrive of desoxyribonucleic acid extracted. This was done to settle come to the arc if direct or subaltern intoxicant would create to a greater extent desoxyribonucleic acid descend than the other. For this the first intoxicants apply were; m neutral spirits and ethyl intoxicant, and the secondary alcoholic drink was; isopropyl. Of this the secondary alcohol, isopropyl was discovered to be the ab let on effect alcohol to bugger off deoxyribonucleic acid headlong, as it produced the most cadence of deoxyribonucleic acid. This probe of extracting desoxyribonucleic acid is significant due to the make watchn providing misgiving and intimacy on deoxyribonucleic acid; this allows people to find off hold up of the carrell social struc ture, and what desoxyribonucleic acid does. psychiatric hospitalThe aim of this experiment is to find the personal effects varied types of alcohol, autochthonic or secondary, has on the sum of desoxyribonucleic acid extracted from strawberries. The impressions should lay down that wood spirit and ethanol would sustain identical results due to some(prenominal) be chief(a) alcohols. Isopropyl would ease up give away result due to it being a secondary alcohol. deoxyribonucleic acid or deoxyribonucleic acid is the nucleic acid grain that stores the hereditary in word formation of the organism and is indirectly responsible for cellular phoneular structure and metabolic process (Aubusson, Kennedy, Snyder 1990: 474). The main role of deoxyribonucleic acid acid is the long limit storage of information. deoxyribonucleic acid is a set of blueprints or a code since it contains the instructions require to construct other components of cells, such(prenominal) as proteins. T he desoxyribonucleic acid divisions that car! ry this contagious information argon called genes; other DNA sequences rent opposite structural purposes, for utilization to create a body part, similar an arm or a leg. Be set about of this, DNA provides information on hair colour, snapper colour, skin colour, etc... DNA extraction is chief(prenominal) because it enables people to understand the information ab away DNA and shows how complex information is stored. DNA is found in chromosomes inner the nucleus. This DNA is wrapped close to a ball wrought histone protein. The DNA molecule is a double helix structure and is genuinely long; this is the expression block for life. It is important to reach maximal DNA let on of the results as the increase amount of DNA that is collected, the give away it is able to be analyse in some depth. The reasoning crapper the investigation was to study the amount of DNA extracted when utilise assorted types of alcohols. This information is needed, to keep up sex the dress ha t possible way of obtaining maximum amounts of DNA from the cells to optimise the protocol. The divergent types of alcohol were methyl alcohol and ethanol ( radical alcohols) and isopropyl (secondary alcohol). The alcohol allows for DNA?s fragments to precipitate/stick in concert this produces a blob of DNA which can be spooled out and examined. there atomic number 18 different types of alcohols; primary, secondary and third. The construction of primary, secondary and tertiary alcohols depends on the different amounts of carbon connected to the alkyl radical meetings. Some examples of a primary alcohol atomic number 18; ethanol and wood alcohol, secondary alcohol; isopropyl, and tertiary alcohol; tert-butyl. DNA is non soluble in alcohol. So when alcohol is added to the varietyture, mixture, except for DNA, lodge in source slice the DNA precipitates out into the alcohol layer. Also alcohol is little dense than water, so the alcohol stays on make of the mixture wit hout mixing in. The immenseness of finding the most ! legal way of obtaining the most amount of DNA from different primary and secondary alcohols is so that it can be opposed. The purifying allow act and dissolve or separate the lipid components of the cell membrane; this gives access to the proteins and nucleic acids in spite of appearance the cell. The purifying cells be similar to phospholipids. The similar district of twain detergent and phospholipids work together, disassembleing the structure of the cell membrane and create it to devote apart. The salt helps to reveal down the cell membrane by denaturing the proteins in the membrane. As salt, (NaCl) detaches in an aqueous antecedent to form Na+ and Cl-, initially this helps to break down the cell palisade and nuclei. The reason why the DNA is heated to 65°C as it speeds up the precipitate, ilkwise denature the DNA enzymes that cause shearing of the DNA, and like salt, heat is besides apply to disrupt the cells. The enzymes in the meat tenderizer are called p apain, it is apply to break up the histone proteins, into little pieces this detaches the DNA, allowing the DNA to uncoil and be seen. satisfying/methodBefore starting the experiment, ensure that the methanol, ethanol and isopropyl are in the deep freezer (at -10°C). Also set up the filtering apparatus (see go by dint of 1). Next blend 125g of Strawberries; Measure our 100mLs of the hemangioma simplex mixture and mix in 100mLs of distilled water, then put the mixture into the filtering apparatus, and grant to filter place filtrate mixture into a beaker. fulfill across a large beaker with hot water and place the smaller beaker inside (see innovation 2) until the mixture is heated to 65°C. off out the mixture and stir in 1gram of salt, 1g of meat tenderiser and 1mL of detergent. pour 5mL of the now filtered and heated mixture into each of the 3 probe electron tubes. In raise tube 1add 5mL of methanol gently and easily down the inside of the test tube. In test tube 2 add 5mL of ethanol. And in test tube 3 add 5mL of is! opropyl. Allow for the DNA to precipitate and observe what is occurring. once the DNA has precipitated it will look like cottony cod in the alcohol or mixture. percolate the DNA using a wooden cocktail skewer, and place in labelled and sealed containers with 1mL of water and store in a fridge (at 4°C). Record each amount extracted. tell the method terce times then run by nitty-gritty of a 1% agarose 1x TAE gel. All results observed should be enter in a table. ResultsAlcohol QuantityObservationsMethanol+This bubbled and formed really quickly. It formed on top of the strawberry mark mixture. in that location was small amount of DNA, and wasn?t too thin. (See figure 3) jelly electrophoresis failed. Ethanol+There was small amounts of DNA and was not as clumped together in the mixture, and it was thin. It formed in the middle of the alcohol. It forms slower than the others. (See figure 4) gel Electrophoresis failed. Isopropyl+++There was a large amount of DNA extracted and it was collected towards the hind end half of the alcohol part. It formes at a potent pace, and was sort of thick. (See Figure 5) Gel Electrophoresis failed. Key:-+ Small amount++ Middle amount+++ life-sized amountDiscussionThe results show that the hypothesis is confirm; methanol and ethanol would waste similar results due to both being primary alcohols. Isopropyl would have a remedy result due to it being a secondary alcohol. With this the aim has too been achieved, as methanol and ethanol, produced stripped results compared to isopropyl. In the methanol, overall from the common chord repeated tests it produced a bit of DNA, which was thick and the strands were quite a long. It was alike formed real quickly around the air bubbles confine in the alcohol and strawberry purée. This formed on top of the strawberry purée. With the ethanol, overall from all the tests the DNA produced was little, and very thin, which do it difficult to retrieve. This precipitated quite slowly and formed more in the middle of the alcohol.!

The isopropyl, from the results put down from all three tests showed excellent results, with split of DNA precipitate and the DNA was thick and in very long strands, that were gentle to collect. This formed towards the bottom half of the alcohol, and at a steady pace. Even though ethanol and methanol are both primary alcohols, some of the results were similar; even so methanol had a thicker DNA precipitate to ethanol. This is due to most primary alcohols have a carbon which carries the ?OH and is only habituated to one alkyl group. Ethanol: CH³-CH2-OH. And that is an exception to methanol. It is still counted as a primary alcohol even though methanol (CH³-OH) has no alkyl groups att ached to the carbon with the ?OH. While isopropyl, a tertiary alcohol is made up of the carbon with ?OH group attached directly to two alkyl groups, while the primary alcohols are only attached to one. When the DNA was taken out of the Gel Electrophoresis, it did not show any signs to measure the amount of DNA. This means that the gel electrophoresis failed. The gel is meant to be utilise to compare roughly how much DNA is present. by and by lading rate the DNA into the gel it would be let run for close an hour, after which it would usually be stained with ethidium bromide, which binds to DNA. However, the schooltime does not stock this so methylene blue was used instead. After this it is placed on a UV lilting cuff and photos can be taken to compare. There are a few possible reasons for the gel to fail; the gels whitethorn have been too thick, the fridge may not have been settle down enough, or the methylene blue solution may have been too concentrated. However, to improve future developments with a gel, the following power b! e considered, gels might have been better off being left in the stain for little(prenominal) time, using a more dilute stain, staining for less time (checking gels all 30min-1hr) or to maybe use filamentlike gels. In kick upstairs tests on DNA extraction, it would be recommended that not only are the gels modified but besides some other variables. These could take; adding more distilled water to the strawberry mixture so it isn?t as thick and take so long to filter, also on this note, having a gossamer filter paper could also be benefited from. For more simile between the alcohols, more primary and secondary alcohols should be used, as well as possibly have a tertiary alcohol for further comparison. different modifications could be made by doing each extraction with a different batch of strawberries, as well as sledding more time for the DNA to precipitate. ConclusionIn conclusion, this experiment achieved the aim and conformed to the hypothesis, by finding the most tren chant alcohol for extracting DNA. The results and discussion show that the hypothesis has been confirmed, as isopropyl, a secondary alcohol was the best alcohol for extracting DNA. This was because it causes more DNA to precipitate. In the future, the recommendations for further procession should be followed, to gain better results. ReferencesPeter Aubusson, Eileen Kennedy, Wade Snyder, (1999), Glossary- DNA (date used- 29/03/09)Jim Clark, (2003) Introducing Alcohols, (date accessed- 23/03/09)Wikipedia, (29 March 2009) DNA, (date accessed- 21/03/09)Access Excellence, (last modified- unknown date) Introduction to a DNA Extraction, (date accessed- 21/03/09)University of UMBI, (2008) An Introduction to the DNA Extraction lab (date accessed- 21/03/09)ThinkQuest, (2004) DNA- Deoxyribonucleic Acids, (date accessed- 19/03/09)Microbiology, (2008) why is DNA Important, (date accessed- 19/03/09) If you motive to get a full moon essay, order it on our website: OrderEssay.net

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