View Single Post
Old 05-15-2017, 04:30 PM   #8
Carcharodon
Member
 
Location: Honolulu, HI

Join Date: Jul 2015
Posts: 40
Default

Quote:
Originally Posted by ymilesz View Post
Hi I am also following the ddRAD protocol from Peterson et al 2012, and I too am having trouble with PCR after size selection. Are you doing the 20uL reactions as suggested as well?
Hi ymilesz,

Sorry for the late response. At first I actually ran 50uL reactions (due to low DNA concentrations in the elute), and then I tried running 25uL reactions.

Interestingly, the ones that tended to fail were the ones that showed higher DNA concentrations post-size-selection. I added water to these to dilute them down to 20ng total DNA content. I'm starting to suspect that this may have affected my results, but I'm not sure.
Carcharodon is offline   Reply With Quote