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Unknown Plasmid Reaction Lab Report

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The molecular lab is a place in which students can perform techniques discussed in lecture. With these techniques one can determine a variety of things within the world of molecular. The main goal for lab this semester was to use different techniques to determine the identity of an unknown plasmid. Early in the semester each student picked an unknown plasmid. Over the course of the next few weeks, the plasmids identity was derived through the results of various techniques. The unknown plasmid selected was plasmid unknown C. Unknown C was subjected to techniques such as DNA isolation, digestions, and gel electrophoreses in order to determine the plasmids identity.The techniques were preformed and the data was collected. The data was analyzed …show more content…
This conclusion was drawn due to the results of different techniques. One technique involved the transformation of E.coli with the unknown plasmid. E.coli was transformed with the unknown and then smeared onto three separate augur plates. Each plate possessed an antibiotic. If growth was seen on the plate, the unknown had a resistance to that antibiotic. The transformed E.coli only colonized the plate which possessed ampicillin. Unknown C therefore has a resistance to the ampicillin antibiotic. Agarose gel electrophoresis technique was preformed to determine the size of the unknown plasmid. The agarose gel was mixed and set in the apparatus. Once set, the unknown plasmid was mixed with some loading buffer and pipetted into a well in the gel. A marker was also placed into a well to serve as a reference. A current was applied and the gel was run. Once completed, it was placed in a UV chamber to view the separation and size. Those results showed that the unknown plasmid was somewhere around five thousand base pairs (Fig1). The list of possible plasmids was narrowed by removing those that were not close in size. The remaining choices were each placed into the NEB cutter program. This

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