Illumina Adapters Thawing/Freezing

[nbafrank]

Hi everyone,

I have been having trouble with getting the protocol to work although it used to in the past. After a little bit of debugging/discussion, I believe that it might be due to bad adapters. So here's my question:

What is the shelf-life of Illumina adapters (in form of ordered oligos and then annealed) with repeated freezing and thawing?

Also is thawing in ice (4C) critical to keep them annealed or RT is fine as well?

Thanks for the advice!

[jlove]

I have found that, after a few freezing and thawing cycles (precise number unknown, maybe 5?), my PCR yields start to decline. So I started aliquotting them out. I store them at -20C and thaw them at 4C for use. I have on occasion left them at RT for a little while (to thaw, for example), but not for long periods of time. I feel superstitious about that, it's probably not the best for annealed linkers.