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Todd McLay 06-19-2014 07:23 PM

Homemade AMPURE beads size selection issue

I made up a batch of the "Serapure" AMPURE beads following the protocol at this website.

The first batch I made worked really well, so I made a second batch and have had some major issues in that the beads no longer do a right-hand side size selection. With the first batch I was using a 0.7-0.8 right to left ratio and getting a nice band between 400-800bp (after adding 50bp of adapter and 100bp of primer), but with the new batch there appears to be no size selection on the right and after PCR I get a smear from 400-1500bp plus.

Has anyone else had a similar issue? I am thinking it might be because of an altered PEG concentration, but I'd like to hear other thoughts.

I'm going to keep the batch to do LHS only selection and make a new batch, but if anyone has any advice on how to mitigate this problem that would be great.


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