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Old 03-22-2011, 02:35 PM   #1
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Location: Monash University, Melbourne, Australia.

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Default Pooling multiplexed libraries after ligation, before PCR

Does anyone do this with the TruSeq kits? Illumina advertised this feature very strongly in their presentations in my area, as a huge time saving feature of the TruSeq kits as compared to the older sample prep kits... yet the product manual doesn't say anything about it. I'm a bit worried about uneven amplification of the libraries during a pooled PCR. Any comments?
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Old 03-24-2011, 03:42 AM   #2
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Would that also cause jumping errors and waste of sequencing reads?
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Old 03-30-2016, 06:41 AM   #3
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Hi I'd like to refresh this thread. Has anyone had success with TruSeq library prep, if you pool your adapter ligated samples prior to the PCR enrichment step?
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Old 03-30-2016, 07:14 AM   #4
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Pooling prior to PCR amplification is not recommended. It precludes the option of adjusting the concentrations of individual libraries to obtain equal read depth.
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Old 03-31-2016, 07:55 AM   #5
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Done once. DO NOT attempt it. Some libraries just dissapeared.
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Old 05-07-2016, 01:10 PM   #6
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We didn't do that. In theory, it may cause unbalanced library pool. Some libraries could have very low sequencing reads.
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