Hi all. I'm constructing libraries for Illumina 100bp paired-end sequencing. I work on a non-model organism, and a large amount of good RNA is difficult to get. Library construction following the TruSeq protocol with Covaris shearing and a size selection step with a gel prior to enrichment has resulted in libraries with very low concentrations - between 4.5ng/uL and 8ng/uL. This is most likely because of the low amount of starting material for the library prep (between 500ng and 3ug). My question is - can I re-enrich this end-product prior to sequencing, or will it create problems?
Any other suggestions would be appreciated.
Thanks!
Any other suggestions would be appreciated.
Thanks!
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