20141009_StudyQuestions_Solved.pdf

I went for the 40 c and 70 c melting points because

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differ by 30°C for the two interactions. I went for the 40 °C and 70 °C melting points because these are both 15 °C away from the given temperature of 55 °C. Many other melting points could have been used in this calculation. This study resource was shared via CourseHero.com
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3. Two restriction enzymes that leave 5’ overhangs are BamHI and BglII. Their recognition sites shown below, where the asterisk indicates the site that is cut: BamHI: 5’ G*GATCC 3’ BglII: 5’ A*GATCT 3’ You perform two separate restriction digests. In one, you digest a plasmid called pBEST with BamHI, which generates one linear molecule (there was only one BamHI site in the circular plasmid). In the other, you digest another plasmid with BglII, which generates two linear molecules (there were two BglII sites in the plasmid). You gel purify the smaller of these two pieces, isolating a fragment that contains Your Favorite Gene (YFG). a. Can you ligate the small piece of DNA containing YFG into the pBEST plasmid? If so, do you need to do anything special? Yes, you can ligate YFG into pBEST. No special steps are required, as BamHI and BglII cut so as to leave compatible ends (both enzymes leave 5’ overhangs with the sequence CTAG (written 5’ to 3’)). b. You have managed to ligate YFG into pBEST to generate pBEST-YFG. Later, you decide to cut the new plasmid to drop YFG out of pBEST-YFG. Indicate below which restriction enzymes you can use to do this: a. BamHI b. BglII c. All of the above d. None of the above Ligation of the BamHI-cut and BglII-cut ends together will either give you the sequence 5’ GGATCT 3’ or 5’ AGATCC 3’. Neither is recognized by either of these two restriction enzymes. This study resource was shared via CourseHero.com
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4. The recognition site for BamHI is G*GATCC while that of PstI is CTGCA*G. You digest a circular plasmid, pMCB, with BamHI (it has 3 BamHI sites) (digest #1). You digest another circular plasmid, p121, with PstI (it has 2 PstI sites) (digest #2). a. How many pieces of DNA do you obtain from each digest? 3 pieces of DNA from digest #1; 2 pieces of DNA from digest #2 b. You purify the smallest piece of DNA from each of the two digestion reactions.
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