How can I tell if I sequenced through the insert? Part 1 | Illumina Video

severe

Library Overamplification with Heteroduplex Formation

High molecular weight fragments appear on Bioanalyzer after PCR due to single-stranded library fragments and heteroduplex formation when PCR primers are depleted. Data quality may be compromised upon sequencing.

💡 4 · ✓ 4

moderate

Excessive Adaptor Dimer Formation

Sharp 127 bp peak visible on Bioanalyzer representing adaptor dimers, reducing the proportion of desired library fragments and overall usable yield.

💡 3 · ✓ 4

moderate

Library Not the Correct Insert Size

Library fragment size distribution is shifted from expected range, with either smaller or larger inserts than intended, or discrepancy between fragment analyzer measurement and sequencer-determined insert size.

💡 5 · ✓ 5