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-   -   Pooling multiplexed libraries after ligation, before PCR (http://seqanswers.com/forums/showthread.php?t=10232)

ScottC 03-22-2011 02:35 PM

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?

chenhongli 03-24-2011 03:42 AM

Would that also cause jumping errors and waste of sequencing reads?

jluk 03-30-2016 06:41 AM

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?

HESmith 03-30-2016 07:14 AM

Pooling prior to PCR amplification is not recommended. It precludes the option of adjusting the concentrations of individual libraries to obtain equal read depth.

Ancestro 03-31-2016 07:55 AM

Done once. DO NOT attempt it. Some libraries just dissapeared.

LOH 05-07-2016 01:10 PM

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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