Qualify Pre-Enrichment Libraries

It is recommended to check the quality or to qualify pre-enrichment libraries before proceeding to enrichment.

If you elect not to check pre-enrichment libraries, perform the following procedure instead to reserve samples for potential troubleshooting later.

1. Transfer 1 μl of each pre-enrichment library to a new 96-well PCR plate.
2. Add 4 µl RSB to each pre-enrichment library.
3. Seal the plate with Microseal 'F' foil seal.
4. Store at -25°C to -15°C for up to 30 days for future troubleshooting if necessary.

Pre-enrichment libraries can be qualified individually, one library at a time, or as a pool before enrichment. Perform the following procedure to qualify pre-enrichment libraries.

1. Assess the quality of 1 µl library or pooled libraries using one of the following methods.
• Add 1 µl RSB to the library, and then analyze the 2 µl volume using the Advanced Analytical Fragment Analyzer with HS-NGS High Sensitivity 474 kit.
• Analyze 1 µl library or pooled libraries using the Agilent Technology 2100 Bioanalyzer using a DNA 1000 kit.

Expect the mean fragment size to be between 300–400 bp and a distribution of DNA fragment size range of 150–1500 bp. The following figures provide example traces.

Example Fragment Analyzer Trace

Fragment Analyzer Trace

Example Bioanalyzer Trace

Bioanalyzer Trace