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Accidentally forgot a base making primers for InFusion (Gibson); Will it work

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#1 Ahrenhase



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Posted 06 March 2014 - 12:34 PM

So my vector PCR product will theoretically have an additional base at the end, where I designed overlapping primers for my insert.  So the exonucleases in the InFusion master mix will chew back the ends of my vector and insert PCR products 5'>3'; however, I'll have an extra base on the vector site.  Will this still work?  I'm hoping the 3'>5' exonuclease activity of my DNA polymerase (Phusion) from PCR will generate some products that are missing this additional base.

Edited by Ahrenhase, 06 March 2014 - 12:34 PM.

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