Lab2_OCON - from the negatively charged pole, or the black...

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Visible band Black electrode or negatively charged pole Open circular DNA Supercoiled DNA Caroline O’Connor Cell Physiology Lab 2 T.A. James Peterson 9/9/09 Title: Bacterial DNA Isolation Lab: Miniprep Isolation of Plasmid DNA 1.) One band was visible on my gel, although the band was not very prominent. Most of the gels run in the lab however did not have visible bands. There are several reasons that the DNA purification may not have worked properly, and consequently may not have had visible bands. First of all, the lysozyme may not have completely bursted open the bacteria, resulting in the DNA and the intracellular contents not being separated. Also, during the resuspension process, the pellet may have been removed along with the remaining liquid, and as a result there would be no nucleic acid pellet. 2.) As mentioned above, there was only one single band visible in the gel. The band moved
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Unformatted text preview: from the negatively charged pole, or the black electrode to the positively charged pole, or the red electrode. This is because DNA has a net negative charge due to its phosphate backbone and therefore will be repelled by a negative charge and attracted by a positive charge. If there were multiple visible bands, however, the smaller DNA pieces would have moved farther than the larger DNA pieces because larger DNA pieces have more difficulty migrating through the pores of the gel. 3.) Supercoiled DNA occurs when circular plasmid DNA becomes wound up. Supercoiled DNA is much smaller and more tightly packed than open circular DNA. Because of its smaller size and density, the supercoiled DNA will migrate farther than the open circular DNA, but it is difficult to predict which band will be larger....
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This note was uploaded on 08/11/2010 for the course IPHY 3060 taught by Professor Allen,davi during the Fall '09 term at Colorado.

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