What if my pass-filter read is < 85 % after sequencing?

  • Updated

When following Illumina guidelines, Tapestri libraries typically generate 85+ % pass-filter reads. If the sequencing data produces fewer than 85 % pass-filter reads, the following sources may have contributed to the suboptimal results:

  • Overclustering/underclustering due to an incorrectly concentrated sequencing library
  • Insufficient PhiX library concentration
  • NaOH (e.g., concentration, prepared on the same day)

Suggested solutions: 

  • Make fresh 5 nM pools of the sample from the saved products after the Library PCR using the Tapestri Sample Quantification Tool (PN 40676). Verify the concentration with Qubit and/or qPCR.
  • Complete the NaOH denaturation step of the library by preparing 0.2 N fresh and incubate for 20 minutes.

Note: If the Q30 scores of Read1, Read2, and the index reads are above 90 %, downstream data processing is recommended.

Share this article:

Was this article helpful?

0 out of 0 found this helpful

Have more questions? Submit a request