DNA Extraction From Chicken Liver

desoxyribonucleic acid stock From Chicken LiverDeoxyribonucleic acid (desoxyribonucleic acid) is the hereditary material in cosmos and almost all other organisms. Nearly every cubicle in a persons body has the same deoxyribonucleic acid. Most DNA is located in the cell nucleus (where it is called nuclear DNA), but a small union of DNA sewer in like manner be found in the mitochondria (where it is called mitochondrial DNA or mtDNA).The in boundation in DNA is stored as a enroll made up of four chemical bases adenine (A), guanine (G), light speed (C), and thymine (T). Human DNA consists of about 3 billion bases, and more(prenominal) than 99 percent of those bases are the same in all people. The order, or sequence, of these bases determines the information avai testing groundle for building and maintaining an organism, similar to the way in which letter of the alphabet appear in a certain order to form words and sentences.DNA bases pair up with each other, A with T and C with G , to form units called base pairs. Each base is also machine-accessible to a sugar molecule and a inorganic inorganic phosphate molecule. Together, a base, sugar, and phosphate are called a nucleotide. Nucleotides are arranged in two longsighted strands that form a spiral called a icon helix. The structure of the double helix is somewhat like a ladder, with the base pairs forming the ladders rungs and the sugar and phosphate molecules forming the vertical sidepieces of the ladder.An important property of DNA is that it can replicate, or make for copies of itself. Each strand of DNA in the double helix can serve as a pattern for duplicating the sequence of bases. This is critical when cells fork because each new cell needs to have an exact reproduction of the DNA present in the old cell.The extraction of DNA from cells and its cultivation are of primary importance to the field of biotechnology and forensics. Extraction and purgation of DNA are the first steps in the analys is and use of goods and services of DNA that allow scientists to detect genetic disorders, produce DNA fingerprints of individuals, and til now create genetically engineered organisms that can produce beneficial products such as insulin, antibiotics, and hormones.Once the DNA has been isolated, it is essential to closely determine its concentration for accompanying manipulation such as cloning or sequence determination.To fix the beat of DNA that extracted by employ spectrophotometry.The aims of this experience is toTo use the properties of DNA to isolate long strands of DNA from liver cells.To determine the yield of DNA isolated from a given kernel of tissue.To examine the light absorb properties of purified DNA.To examne the relationship between the concentration of a DNA issue and the absorbnce at 595nm of DNA-diphenylamine solution.To generate a standrad curve relating DNA concentraton with the absorbance of DNA-diphenylamine solutions.To use a archetype curve to dete rmine the concentration of an unknown DNA solution.Materials and MethodsAs per lab manual.ResultsFirstly, the chicken liver cell homogenate is treated with a salt solution such as NaCl and a detergent solution containing the compound SDS (sodiumdodecyl sulfate). These solutions become down and emulsify the fat proteins that make up a cell membrane. Finally, ethanol is added because DNA is soluble in water. After adding ethanol a relatively clear aqueous will be produced, the first mould is the milky solution that is the aqueous phase with DNA, the middle layer is the unscathed (precipitate proteins). The bottom layer is a clear solution (organic). The DNA can be spooled (wound) on a stirring rod and pulled from the solution at this point. The fare of DNA solution we got is 5.4ml.Than we put the DNA solution in 2ml tube (1.041g).The total weight of DNA solution and tube is 1.106g. The amount of DNA we got is 1.106-1.041g = 0.065g.Next we prepare 4 measuring rod tubes by adding TE buffer (ml) to the DNA standard solution (ml). And also added to each of the 3 samples of my DNA. The total DNA (mg) is enter in the remand 1. The observed colorize change of 4 standard tube and my 3 samples are recorded in table 2 and 3. We pipette the DNA samples and each standards tubes into dissolve wells of a 96 well microtitre plate. We measured the absorbance at 595nm of the DNA-diphenylamine solutions using the plate reader. Our results are shown in the graph with the used of the reading of table 4. Form the graph we find that the concentration of undiluted DNA is 0.232=0.46mg/ml. discussion and ConclusionsFor this sample we determinate the yield of the DNA isolate from given amount of tissue is1g - 63mg0.065g - 4.095mg (wet weight of the DNA to dry weight)3ml - 4.095mg5.4ml - 7.371mg (DNA in the entire aqueous phase is collected)3. 4ml - 7.371mg5.3ml - 9.767mgThe final calculation of the dry DNA is 9.767mg/g liver.For the experiment we examine that the light absorb ing properties of purified DNA. The wavelength is range 220-300nm. The wavelength of the DNA is 260nm.We also careful that the yield of DNA per g of liver from Lab 2 isThe amount (mg) of DNA contain = 0.461.5=0.69mgAqueous from lab 1 = 5.4mg0.69/2 =0.345mg(0.3455.4)/3 = 0.621mgThe final value in mg of dry DNA/g liver is 0.621mg/g.In the end of the experiments, we managed to complete our objectives. In summary, we learn that the alcoholic beverage can causes DNA to precipitate, or settle out of the solution, leaving behind all the cellular components that arent soluble in alcohol. As alcohol is little dense than water, so it floats on top forming two separate layers. We also learn that the usefulness of spectrophotometry is that diphenylamine only reacts with DNA more accurate as RNA would not be determined. The disadvantage of spectrophotometry is that it always requires standard solution. The advantage of calculating of yield by its weight is that it does not require standard s olution. The disadvantage of calculating of yield by its weight is that it is less accurate as RNA is counted in.