Plasmid Isolation And Its Effects On The Dna Essay

1208 Words Oct 15th, 2016 5 Pages
Our goal within the plasmid isolation lab was to isolate the bacterial plasmid without any damage to the DNA. The formation of the clear lysate was essential because it allowed us to isolate the bacterial cells from removed media and later expose the DNA. Resuspension solution helped to resuspend the cells, after centrifugation, and prepare the cells for lysis solution. The lysis solution functions to degrade the cell membrane and spill its components. Then the Alkaline Protease functioned as the inhibitor of any DNases and denatured dsDNA to ssDNA shortly, by disturbing H-bonds between bases. Finally, the Neutralizing Solution was helpful into stabilizing the pH, from a basic solution formed by the lysis and protease solution.
The DNA was now ready to be isolated from the reset of its components with the use of a spin column. The spin column contains a positively charged filtered system that allows the DNA to bind to it while filtering anything else. To ensure the filtration of any unwanted material, anything other than DNA, the DNA was washed several times with ethanol. Ethanol changes the DNA structure so that the DNA molecules aggregate and precipitate from the solution. However, since most salts and organic molecules are soluble in 70% ethanol, they stay in solution and precipitated DNA can be separated from them by centrifugation. After washing with ethanol several times, assuming that DNA is now only present in the spin column, we need to dry wash the column to…

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