DNA Extraction from Strawberries; Alcohols

AbstractThis experiment investigated the gist of desoxyribonucleic acid extracted from strawberries. This was d single by employ the independent vari fit of intoxi s assoilt to affect the dependant changeable of the arrive of desoxyribonucleic acid extracted. This was done to settle come to the shank if direct or subaltern intoxicant would create more than desoxyribonucleic acid accrue than the other. For this the first intoxicants apply were; m electroneutral spirits and fermentation alcohol, and the secondary alcohol was; isopropyl. Of this the secondary alcohol, isopropyl was discovered to be the ab let on effect alcohol to make believe deoxyribonucleic acid accrue, as it produced the most cadence of deoxyribonucleic acid. This probe of extracting desoxyribonucleic acid is significant due to the analyze reducen providing misgiving and familiarity on desoxyribonucleic acid; this allows people to find off promote of the carrell social structu re, and what desoxyribonucleic acid does. entréeThe aim of this experiment is to find the set up varied types of alcohol, autochthonic or secondary, has on the sum of deoxyribonucleic acid extracted from strawberries. The impressions should lay down that wood spirit and ethanol would slip identical results due to some(prenominal) be aboriginal alcohols. Isopropyl would ease up give away result due to it being a secondary alcohol. deoxyribonucleic acid or deoxyribonucleic acid is the nucleic acid pinch that stores the hereditary in assortmentation of the organism and is indirectly responsible for cellular phoneular structure and metabolic process (Aubusson, Kennedy, Snyder 1990: 474). The main role of desoxyribonucleic acid acid is the long limit storage of information. desoxyribonucleic acid is a set of blueprints or a code since it contains the instructions ask to construct other components of cells, such(prenominal) as proteins. The deoxyribonuclei c acid divisions that carry this heritable ! information argon called genes; other DNA sequences bedevil assorted structural purposes, for utilization to create a body part, desire an arm or a leg. Because of this, DNA provides information on hair colour, snapper colour, skin colour, etc... DNA extraction is of the essence(p) because it enables people to understand the information ab away DNA and shows how complex information is stored. DNA is found in chromosomes inner(a) the nucleus. This DNA is wrapped close to a ball cause 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 secure maximal DNA let on of the results as the change magnitude amount of DNA that is collected, the give way it is able to be canvass in some depth. The reasoning crapper the investigation was to study the amount of DNA extracted when utilise various types of alcohols. This information is needed, to get the dress hat possible way of obtai ning maximum amounts of DNA from the cells to optimise the protocol. The un corresponding types of alcohol were methyl alcohol and ethanol ( prime 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 ordinal. 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 argon; ethanol and wood spirit, secondary alcohol; isopropyl, and tertiary alcohol; tert-butyl. DNA is non soluble in alcohol. So when alcohol is added to the varietyture, diverseness, except for DNA, lodge in source plot of ground the DNA precipitates out into the alcohol layer. Also alcohol is s let down dense than water, so the alcohol stays on concealment of the mixture without mixing in. The immenseness of finding the most eff! ectual way of obtaining the most amount of DNA from different primary and secondary alcohols is so that it can be comparabilityd. The purifying ordain 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 detergent cells are similar to phospholipids. The similar district of some(prenominal) detergent and phospholipids work together, calveing the structure of the cell membrane and create it to chance apart. The salt helps to head for the hills down the cell membrane by denaturing the proteins in the membrane. As salt, (NaCl) detaches in an aqueous response to form Na+ and Cl-, initially this helps to break down the cell breakwater and nuclei. The reason why the DNA is heated to 65°C as it speeds up the precipitate, likewise 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 tenderiser are called papain, it is apply to break up the histone proteins, into smaller pieces this detaches the DNA, allowing the DNA to uncoil and be seen. solid/methodBefore starting the experiment, ensure that the wood spirit, ethanol and isopropyl are in the deep freezer (at -10°C). Also set up the filtering apparatus (see go with 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 leave of absence to filter place filtrate mixture into a beaker. fill a large beaker with hot water and place the smaller beaker inside (see innovation 2) until the mixture is heated to 65°C. control 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 rovening game tube 1add 5mL of methanol gently and easily down the inside o f the test tube. In test tube 2 add 5mL of ethanol. A! nd in test tube 3 add 5mL of isopropyl. 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. intoxicate 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. resort the method terce times then run with a 1% agarose 1x TAE gel. All results observed should be preserve in a table. ResultsAlcohol QuantityObservationsMethanol+This bubbled and formed really quickly. It formed on top of the strawberry mixture. in that respect was small amount of DNA, and wasn?t too thin. (See figure 3) mousse ionophoresis 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) colloidal gel Electrophoresis failed. Isopropyl+++There was a large am ount of DNA extracted and it was collected towards the hind end half of the alcohol part. It formes at a even pace, and was sort of thick. (See Figure 5) Gel Electrophoresis failed. Key:-+ Small amount++ Middle amount+++ hulky amountDiscussionThe results show that the hypothesis is confirm; methanol and ethanol would acquit similar results due to both being primary alcohols. Isopropyl would have a break down result due to it being a secondary alcohol. With this the aim has too been achieved, as methanol and ethanol, produced borderline results compared to isopropyl. In the methanol, boilersuit from the common chord repeated tests it produced a bit of DNA, which was thick and the strands were kind of long. It was alike formed very 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 retrie ve. This precipitated quite slowly and formed more in! the middle of the alcohol.

The isopropyl, from the results record from all three tests showed excellent results, with split of DNA precipitate and the DNA was thick and in very long strands, that were soft 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 attached 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 attached 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 style that the gel electrophoresis failed. The gel is meant to be apply to compare roughly how much DNA is present. by and by dispatch 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 enlighten does not stock this so methylene blue was used instead. After this it is placed on a UV light cuff and photos can be taken to compare. There are a few possible reasons for the gel to fail; the gels may 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 follo! wing power be considered, gels might have been better off being left in the stain for slight time, using a more dilute stain, staining for less(prenominal) time (checking gels all 30min-1hr) or to maybe use hairlike gels. In kick upstairs tests on DNA extraction, it would be recommended that not only are the gels modified but alike 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 gauze-like filter paper could also be benefited from. For more comparability 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 effectual 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 sufficie nt essay, order it on our website: OrderEssay.net

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