Experiment 4 - 1 Chris Stanton April 16, 2009 Leah,...

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1 Chris Stanton April 16, 2009 Leah, Thursday Rm 41 Experiment 4: Analysis of Exconjugants by PCR and Gel Electrophoresis Gel electrophoresis in this experiment showed different DNA fragments in the E.coli CSH55 (an F- strain), CSH61 (an Hfr strain), and an exconjugant. The gel that ours was run, unfortunately, must have not been loaded correctly because we were unable to read our results. The make up sample gel we obtained after that had correct data and was loaded into Gel A, Section 1. From left to right, the lanes consisted of CSH61 with Primer 5, the Exconjugant with Primer 5, CSH55 with Primer 5, CSH61 with Primer 8, the Exconjugant with Primer 8, and CSH55 with Primer 8. See the attached photocopy of the gel. The DNA fragment amplified by Primer 8 was about 800 bp long and was only observed in the F- strain. The DNA fragment amplified by Primer 5 was about 460 bp long and was observed in both the Hfr strain and in the exconjugant. Using the partial sequence of the E. coli genome provided, the expected size of the PCR fragment with Primer pair 5 was 473 bases. The expected size of DNA using Primer pair 8 was 115358 bases, or about 115 kb. It is known that this PCR could not amplify a segment larger than 4 kb; therefore, it is assumed that in wild-type DNA no PCR product will form using Primer pair 8. The PCR for the CSH55 strain and CSH61 strain served as controls with which
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Experiment 4 - 1 Chris Stanton April 16, 2009 Leah,...

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