EMSA with PCR fragments

trinity84 · #1 (**permalink**) 05-23-2008, 04:46 PM

Hi,

Has anyone tried doing EMSA with PCR fragments of a promoter, instead of oligos?

Thanks!
T84

gandalfthegrey · #2 (**permalink**) 09-04-2008, 10:29 PM

Hi, yes this is possible however you must label your PCR products after PCR. This is not easily done however if you digest or somehow alter the DNA to have ends restriction digested you can easily Klenow in p32-Atp or other nucleotide in. Also, its been known to be done with biotin.

There was a method to biotinylate DNA fragments even undigested after PCR but I forget now its been a while... I think it used a special enzyme.

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EMSA with PCR fragments

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