The sparQ DNA Frag and Library Prep kit optimises the integration of enzymatic fragmentation into a two-step protocol for the streamlined construction of libraries for sequencing on Illumina® NGS platforms. A single-tube enzyme mix facilitates the combination of fragmentation and DNA polishing minimising DNA over fragmentation while greatly simplifying library.

• Simple two step workflow employs a unique enzyme mix safeguarding samples from over fragmentation
• Robust enzymatic fragmentation and consistent results
• Flexible generation of high quality libraries from 1 ng to 1 µg of input DNA
• PCR-free workflows enabled from 100 ng
• Minimised bias across challenging regions for improved sequencing results

The streamlined workflow can be completed in under 3 hours with minimal hands-on time. Designed to combine the highest efficiency enzymes for fragmentation, polishing, adapter ligation and amplification, the sparQ enzymes generate superior yields of sequencing ready libraries over a broad range of input DNA.

Double-stranded DNA molecules are fragmented followed by DNA polishing reactions where 5'-phosphorylated and 3'-dA-tailed DNA fragments suitable for direct ligation of sequencing adapters are generated without the need for an intervening cleanup step. The HiFi PCR master mix and primer mix allow unbiased amplification of fragments with appropriate adapters ligated to both ends.

The highly efficient library prep reduces coverage bias resulting in superior quality needed and expected, to minimise coverage gaps especially for challenging regions like GC- and AT-rich sequences. Reproducible and uniform genome coverage is achieved independent of input DNA amounts and closely resembles coverage obtained using mechanical shearing workflows resulting in optimised sequencing outcomes.