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#1
02-28-2010, 08:12 PM
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 Overlap PCR failing to introduce mutations Hi, I attempted to use Overlap PCR to introduce two mutations (F to A mutation) into the DNA sequence of a protein im studying. The mutations were carried on two seperate primers i.e. one on primer B and the second on primer C. PCR products AB and CD were the correct size when checked on a gel. Again the final product was the expected size. However after sending the final PCR product away for sequencing, the results i recieved back indicated that no mutations had been introduced and after a blast was proformed it was a 100% identical to the wild-type proteins DNA sequence. Can anyone help me understand why that happened?  and what i can do troubleshoot this?would appreciate it  |
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#2
03-02-2010, 08:50 AM
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 Re: Overlap PCR failing to introduce mutations I can't really help you with troubleshooting, but I recommend trying the "QuikChange Site-Directed Mutagenesis Kit" by Stratagene. It has never failed me! |
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| failing , help with overlap pcr , introduce , mutations , no mutations via pcr , overlap , overlap pcr , pcr |
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