Question

Review. Consider how one might label the 5'ends of DNA for a footprinting experiment where you visualize where on the DNA a transacting protein binds to the cis acting promoter sequence The 5 phosphate(s) at the S'ends a DNA can be removed using a phosphatase, like calf intestinal phosphatase (CIP), in vitro, leaving a 5-hydroxyl group at the end of DNA CIP has no impact on the phosphates along the backbone of the DNA, just the 5' end phosphate and of course, there is no phosphate at the end of DNA. This phosphatase is effectively denatured by heating at 75C for 10 minutes. DNA will remain base paired at 75C. So, you can treat the DNA with CIP, then heat the solution to 75C for 10 minutes, and now you have a solution of 5'dephosphorylated DNA and "dead" CIP - the denatured CIP makes no impact in any future reactions with the 5 dephosphorylated DNA. T4 polynucleotide kainase (T4 PNK) is an enzyme coded for in the pervome of the T4 bacteriophage. In vitro, the T4 PNK can transfer an organic phosphate from the gamma position on ATP to a 5-hydroxyl group of DNA or RNA like the DNA you Which of the following radioactively labeled ATP's would you correctly use to label CIP treated DNA in the presence of T4 PNK? (NOTE: the red star indicates the radioactive atorn он он ON -O-P-O-P-O- AHCO Онон a H50-PHOT OH OH OCHOPOP-DP-0 OM Мас

Answer 1

Option 3

T4 polynucleotide kinase transfer gamma po4 to 5'end of DNA . Since the above option has radiolabled po4 at gamma terminal position, this will be used

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