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Old 10-16-2013, 02:52 PM   #1
lescagasan
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Default PCR based test for TruSeq library contamination in Pre-PCR lab?

Hi,
I am looking for a simple PCR based assay to evaluate if there is TruSeq amplified library present in our Pre-PCR lab. I would like to use the Bioanalyzer to quantitate. Would something as simple as designing primers to the P5/P7 and amplifying work? Could I just use the leftover primers from the library prep kit? How many cycles would you recommend? Any input or ideas welcomed.
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Old 12-14-2013, 01:04 PM   #2
Genohub
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Your pre-PCR libraries probably contain doubly ligated product which in itself can amplify. How about running qPCR with some PCR amplified positive controls and comparing Ct values. I can't think of a good way for you to do with with a bioanalyzer. Are you having contamination issues?

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Old 12-21-2013, 09:06 AM   #3
ECO
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I use Kapa's qPCR quantitation kit (which as you said is P5/P7 primers). Bioanalyzer won't be sensitive enough.
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