Hi,
Is there any obvious explanation why there would be a lot (~50%) of poly-A (or Poly-T, since this was bidirectional lib prep) in the output ?
RNA was purified using Poly-A+ enrichment (purified twice). Is it possible that the library was overamplified, hence the "PCR-winners" (polyA/T have lower GC) ? Is the library usable and the only drawback would be a lot of sequencing wasted on poly-A ?
Is it possible that the RNA was already digested, hence the only fragment retained in enrichment were the poly-A them self?
Thanks!
Is there any obvious explanation why there would be a lot (~50%) of poly-A (or Poly-T, since this was bidirectional lib prep) in the output ?
RNA was purified using Poly-A+ enrichment (purified twice). Is it possible that the library was overamplified, hence the "PCR-winners" (polyA/T have lower GC) ? Is the library usable and the only drawback would be a lot of sequencing wasted on poly-A ?
Is it possible that the RNA was already digested, hence the only fragment retained in enrichment were the poly-A them self?
Thanks!
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