Targeted sgRNA NGS for Pooled CRISPR Libraries (Hybridization Capture-based)

FLI-Seq™ amplifies only the CRISPR insert, avoiding PCR artefacts

While performing CRISPR screens, researchers face challenges with the PCR amplification step before sequencing. The large amount of DNA used in published protocols inhibits the PCR reaction. To get enough DNA for sequencing, scientists have to increase the number of PCR cycles. Increased PCR cycles can be problematic when amplifying a library, as the final results are usually PCR duplicates or artifacts with no useful information.

FLI-Seq™ (Fast Library Insert – Sequencing) solves this problem by removing undesired genomic DNA while enriching for the region of interest. In the case of CRISPR screens it will enrich the DNA fragment containing the single guide RNA (sgRNA).

Once enriched, this fragment can be easily amplified with less PCR cycles, avoiding PCR duplications and obtaining robust and reproducible results.