Hello,
I'm preparing ~250 individually barcoded samples (16s metagenomics) that will be pooled in equimolar quantities for a lane of MiSeq.
I am planning to pool 25ng of each sample; however most protocols that I've read use much more than this.
Are there any disadvantages to pooling smaller quantities of PCR product from each sample?
It seems to me that as long as the final pooled library can be submitted at 10nM then all should be ok, but I could use a second opinion.
Thanks for you help!
I'm preparing ~250 individually barcoded samples (16s metagenomics) that will be pooled in equimolar quantities for a lane of MiSeq.
I am planning to pool 25ng of each sample; however most protocols that I've read use much more than this.
Are there any disadvantages to pooling smaller quantities of PCR product from each sample?
It seems to me that as long as the final pooled library can be submitted at 10nM then all should be ok, but I could use a second opinion.
Thanks for you help!
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