Attach your TABLE (Provided above showing the migration distances for the MW mar
ID: 168289 • Letter: A
Question
Attach your TABLE (Provided above showing the migration distances for the MW markers (1 kb ladder), PCR fragment, and vector, both digested and undigested. You should have calculated the sizes for each band you detected on your gel using the standard curve you generated from the MWM). Make sure these sizes are entered on the TABLE. Attach your log-linear graph with standard curve 1. How many bands did you detect in the lane with undigested plasmid? What size(s) were they? Explain each band. 2. How many bands did you detect in the plasmid cut with BamHI? Basing your answer on this result, how many sites are there in the plasmid? 3. Compare the insert digested with BamHI with the undigested insert: do you see a size difference? Explain. 4. Given the sizes of the vector and PCR product, what is the expected size of the plasmid constructed by ligating the two? Explain.Explanation / Answer
1.2 bands. They are due to super coiled (1650kb) and nicked circles (12000kb) of plasmid. Highly supercoiled DNA moves longer distance upto 4.8cm.
2. 2 Bands. The 2 Bam HI sites are present in plasmids. After digest with Bam HI the original plasmid DNA become
linear 12000Kb and the undigested DNA goes to 4.8 cm due to supercoil. The 2 restriction site for BamHI gives a
2000bp of insert.
3.The PCR fragment (undigest insert) is digested with Bam HI it will remove some of DNA base pairs present in the PCR fragment. The Digest DNA (insert digest) contains less bp compare to undigest insert.
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