Primers for QuikChange - desalted vs PAGE purified

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Primers for QuikChange - desalted vs PAGE purified

Post by seanyumcloughlin » Sun Sep 02, 2007 11:16 pm

Hi Everyone,

I'm new here, nice site, nice to be here.

I'm in a new lab where they use short (30 nt), desalted primers to do the QuikChange rxn. Stratagene recommmends HPLC or PAGE purified primers. Using ordinary primers, I get a nice yield of PCR product, but very few transformants. I used to use PAGE purified primers and got more transformants. The transformation efficiency of the cells is high (10 to the 10). Seems the PCR product is not propagated well. Any ideas?

Sorry if this topic has been discussed previously.



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Post by ShineGene » Sun Sep 23, 2007 2:15 am

You had better use Page pruified is better to desalted primer.

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Post by Ammie » Wed Oct 03, 2007 11:08 am

Hi! I used desalted primers for multi QuikChange, and out of three sequenced colonies I got one with four of five mutations and the other with three of five. My primers were all 40nt long.

Maybe it would have worked even better with page-purifyed primers.

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