DNA extraction is an essential technique used in many scientific research endeavours. In my gene and genome analysis lab, BIO383H, we extracted DNA from a young Brachypodium distachyon leaf. DNA is found in the nucleus of cells. As such, we had to disrupt cells and tissues to obtain the DNA. First, we did that by mechanically grinding the leaves. Then we had to use detergent, found in the DNA extraction buffer, to dissolve the solubilize the plasma membrane and other membranes within the cell. Within the extraction buffer is also a metal chelating compund which locks away calcium and magnesium ions, allowing the denaturation of proteins. High temperature was also used to augment the efficiency of extraction buffer and hasten cellular disintegration. After breaking up cells and tissues, we had to prevent the degradation of DNA by DNAse enzymes. The metal chelating compound already helped us with this by locking away calcium and magnesium ions, which are essential co-factors for enzyme activity. Next, we had to remove undesirable products. We did this by rounds of centrifugation and then precipitating out the DNA by using high levels of sodium and isopropanol. To truly purify the DNA, 70% ethanol was used to wash it. Then it was dissolved in T10E1 before being stored in a freezer.
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