Could have been improved with a more thorough mixture

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could have been improved with a more thorough mixture of dye throughout solution, a lower vo Fig. 1. Confirmation of YMP met mutants using gel electrophoresis. The met2, met3, and met17 auxotrophs were mixed with 6X loading dye (bromophenol blue, xylene cyanol, and glycerol) and analyzed on an agarose gel in TAE buffer. The 1.25% agarose gel ran for 20 minutes at 120V. Amplified DNA was stained with 0.05mg/mL ethidium bromide (EtBr) and visualized with a UV Transilluminator. A. Standard Ladder: Lambda EcoRI + Hind III, showed multiple bands with 21226, 5148, 4973, 4268, 3530, 2027, 1904, 1584 bp. B. Genomic DNA from YMP19 with gene-specific primer, MET3 Primer A, and KAN Primer B. No band was visible. C. Genomic DNA from YMP19 with gene-specific primer, MET25 Primer A, and KAN Primer B. A 1700 bp band formed. D.
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Genomic DNA from YMP12 with gene-specific primer, MET2 Primer A, and KAN Primer B. No band was visible. E. Genomic DNA from YMP12 with a gene-specific primer, MET3 Primer A, and KAN Primer B. A 1904 bp band formed. F. Genomic DNA from YMP24 with a gene-specific primer, MET17 Primer A, and KAN Primer B. A band was not visible under UV light. G. Genomic DNA from YMP24 with a gene-specific primer, MET2 Primer A, and KAN Primer B. A 1904 bp band was visible under UV light. To further confirm the identity of mutated strains, YMP12 for example, I would perform a pair of PCR amplifications to modify the reaction between MET3 Primer A and KAN Primer B. In one
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Christopher Reinemann
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