Question

6a. What is the size of the uncut pKAN? __________________ What would this look like if you ran it on a gel? Explain.

6b. If you cut pKAN with one enzyme, how many fragments would you see if you ran the sample on gel electrophoresis. What size would the fragment(s) be?

6c. If you performed a double digest of pKAN with BamHI and HindIII, how many fragments would you see if you ran the sample on gel electrophoresis? What size would the fragment(s) be (indicate them from smallest to largest)?

Answer 1

6a) size of uncut plasmid will be less than 5.5kb which is actual size but due to circular in nature it actual size become less and band will form around 3kb

6b) pKAN size is 4194 kb which become linear after digesting with single enzyme

6c) if we digest pKAN by BamHI and HindIII then it will cut at 2095bp and 234bp from start position of plasmid then the fragment size will be 2095bp-234bp=1861bp

While remaining fragment of plasmid will be 4194-1861=2333bp

So on gel electrophoresis two fragments will be observed one 2333bp and another 1861bp.

Hope it's clear.. thanks