Using Information Obtained From The Single And Double Digests

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This experiment overall was very successful as using information obtained from the results of both the single and double digests, a credible restriction map for the unknown plasmid could be constructed. Within this experiment, both single digest and double digests consisting of three restriction endonucleases were used in order to map out the restriction sites of the enzymes making up an unknown plasmid. In order to separate the DNA fragments by their distinct number of base pairs, it was necessary to run an agarose gel electrophoresis. For this particular experiment, a 1% agarose gel was used as this concentration ultimately results in pores that can separate the DNA by size. The process of gel electrophoresis is made possible by the electric current that is used to move the samples of DNA throughout the gel. For this particular experiment, the gel was run at a current of 100 volts. As a result of the phosphate backbone of DNA, DNA is negatively charged. Because DNA is negatively charged, it moves away from the negative electrode and moves down the gel toward the positive electrode as it is attracted that way. What allows for the resistance the fragments face when moving away from the negative electrode is the texture of the gel itself. As a result, the slower fragments of DNA have the ability to move at a faster rate than the larger fragments of DNA. Each sample loaded into the gel contains a loading-dye for two reasons. One reason is that the loading dye contains
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