Animal Genomic Dna Extraction Essay

759 words - 4 pages

For the 2C experiment, we have been using chicken’s muscle tissue sample to obtain the DNA sample. The muscle’s sample had been minced in order to form a suspension of cell. There is another method that can be used to disperse the tissue that is by pouring liquid nitrogen into the muscle’s tissue and grind it by using whether pestle and mortar or a plastic grinder. If the sample tissue is not too large, a homogenizer can be used. Using large amount of tissue is not recommended as it could give poor DNA quality and a worse yield.
Before the tissue being minced, it has been washed by using phosphate buffer saline (PBS) solution. This solution is isotonic and non-toxic to the cells and has the ...view middle of the document...

For this experiment, we have used a Phenol: Chloroform solution to this step. Both of phenol and chloroform have the same general attributes. Phenol usually is a buffer solution that comes with two layers. The top layer is the excess buffer while the bottom layer is the saturated phenol. Every time the sample was being centrifuged, there will be two layers that can be spotted in the tubes. The upper layer will contains the DNA while the bottom layer will contains lipids, proteins and polysaccharides of the cells. Even though chloroform’s attribution is same as phenol, it also could stabilize the unstable boundary between the aqueous and the organic layers.
For each centrifugation step, the sample will get cleaner and cleaner. The last step which is the DNA precipitation process, we have used sodium acetate (NaOAc). This NaOAc solution is widely used in this process as it can keep a relatively constant pH level. NaOAc is also a type of salt that could neutralize the negative charge of the DNA (Sigma-Aldrich, 2014b). When the solution has been centrifuged, again, a pellet of DNA could be seen. The pellet later will be washed with 70% of ethanol to remove the excess...

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