Originally posted by Dynamac
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Getting the enrichment down to 5-10% will probably help dramatically with the passing filter. I know of one lab that won't sequence amplicons that enrich over 7%! High enrichment leads to high numbers of mixed beads which leads to high percentage of too short quality, and several of the filters are included in the too short quality category, which makes it even more confusing!
Also, are you reducing the amplification primer? I had one set of amplicons that I was seeing the same problem with, except more extreme. There were no CATG control beads showing up at all. Turns out that the samples were amplifying so well that the signal drowned out the CATG control beads.
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Originally posted by Anthony.287 View PostGetting the enrichment down to 5-10% will probably help dramatically with the passing filter. I know of one lab that won't sequence amplicons that enrich over 7%! High enrichment leads to high numbers of mixed beads which leads to high percentage of too short quality, and several of the filters are included in the too short quality category, which makes it even more confusing!
Also, are you reducing the amplification primer? I had one set of amplicons that I was seeing the same problem with, except more extreme. There were no CATG control beads showing up at all. Turns out that the samples were amplifying so well that the signal drowned out the CATG control beads.
Thanks!
J
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