- Optimized Library Prep
- Fewer PCR Reactions and Cycles Required
- Highly Reproducible sgRNA Results
- Less Sequencing Coverage Needed
While performing CRISPR screens, researchers usually face problems with the PCR amplification step before sequencing.
The very large amount of DNA used in all published protocols inhibits the PCR, so to get enough DNA for sequencing, scientists have to increase the number of PCR cycles. Increased PCR cycles is a problem when amplifying a library, as the final results are usually PCR duplicates with no “useful” information.
FLI-Seq™ 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.
Each Kit includes:
- 3× FLI-Seq Buffer
- FLI-Seq Probes
- FLI-Seq Beads
- sgDNA Primers Mix
- 2× PCR Mix
- Bead Elution Buffer
- Indexed Primers