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)
- 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.