Monday, December 5, 2016

Human DNA Extraction Lab

        In this lab, we asked how DNA can be separated from cheek cells to study. We found that significant amounts of DNA can be extracted as a visible precipitate. The thread-like structure could be seen suspended in the isopropanol alcohol, after the layer of nonpolar alcohol was added on top of the solution with DNA. It has been proven that through homogenization, lysis, and precipitation, one can extract DNA. This is done through breaking down the membranes and nuclear material with polar liquid, lysing the membranes, emulsifying the proteins and lipids, and breaking down the histones in the DNA. Then, by adding a polar substance as a layer, the DNA will fall out of solution at the interface and become a precipitate. Our data supports this claim and is the result of such a process, using salt as a precipation facilitator, detergent as a disruptor, and pineapple juice as a catabolic protease to help break down different structures in the cell and free the DNA.
DNA precipitates into the ispropanol alcohol
        Some errors people in our lab group made led to alcohol and solution mixing, thus precluding the DNA from becoming a precipitate. In one experiment, the inversion to mix detergent with the solution was done too quickly and violently, thus leading to bubbles forming and disrupting the liquid surface. This would have kept the DNA from successfully precipitating at the interface, and could have mixed up or damaged the DNA strands in the solution. Another error was not measuring very precisely, which lead to a variety of results, some unsuccessful. We simply estimated amounts (e.g. pinch of salt), which may have led to imprecise measurement and thus a failed extraction with a certain substance being too dilute or concentrated. In a future experiment, I would suggest making sure to mix slowly, which would keep the solution from forming many bubbles. Additionally, keeping measurements precise, such as using measuring spoons and graduated cylinders, would give less variation and a higher success rate in general.
Precipitation is unsuccessful due to violent inversion
        This lab was done to demonstrate how DNA can be extracted from inside the nucleus of a cell. Moreover, I also learned how the membranes, histones, and nuclear material must all be broken down before the DNA can be extracted. This helped me understand how much protection the DNA has, due to it holding the genetic information. Not only does it never leave the nucleus (mRNA copies and leaves instead), there are many layers of membranes to keep it from being damaged by substances outside the cell. Outside of these concepts, based on my experience from this lab, I also now know how one might break down cell membranes to access, not only the DNA, but any organelle one might be studying using the techniques of homogenization, lysis, and precipitation.

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