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Old 07-08-2013, 07:16 AM   #1
Location: Atlanta

Join Date: Dec 2012
Posts: 70
Default How long the template can be stored at -20?

Dear everyone.

I have diluted my libraries to 12pM templates about 2 weeks ago ,and stored at -20 degree. Can I use those templates for cluster generation today? Or I must use fresh diluted templates?

Thank you so much!
illuminaGA is offline   Reply With Quote
Old 07-08-2013, 07:46 AM   #2
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Location: London

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We've re-run 12pM a couple of times before after 2 weeks. The data was fine but we did see a marginal reduction in cluster density. We don't do it often enough to say with certainty that it'd be OK though.
We've kept 20pM for 2 months and that seemed to be fine too. In fact we now use a 20pM stock to prepare our PhiX MiSeq spike-ins and just make up some fresh 20pM every few weeks.
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Old 07-17-2013, 10:52 AM   #3
Location: UK

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Based on TruSeq library, You absolutely have to run it the same day after multiplexing is finished. This will ensure ~800k/mm2 clusters. We noticed that running the very same sample (stored on SGP, non-diluted) after just 4 days results in a drop from 850k to ~500k. After two weeks we had results ~350k/mm2...Im not sure if there is anything that we can do - overloading works up to a limit of NaOH concentration.
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