Help with PCR reaction Phusion Flash - wrong sized fragment

L Celeste via methods%40net.bio.net (by celesteel from gmail.com)
Tue May 26 18:01:24 EST 2009


1. When we did sequencing, we only send the DNA template as well as the
primer, we didn't actually sequence the 2 kb fragment (and it actually is
more like 2.5 kb). So our assumption is that that 2 kb fragment we are
seeing constantly does contain the 1 kb from the sequencing reaction.

2. What number of cycles would you suggest? I have tried 25, 27 and 30
cycles.

Thanks


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