A new high fidelity PCR master mix with broad NGS workflow compatibility and superior performance in high sensitivity applications

The widespread adoption of sensitive mutation profiling in precision oncology and the emergence of next-generation sequencing platforms with increasing data quality demand high accuracy library preparation tools. Additionally, PCR master mixes used in NGS library prep are often specialized for specific applications, sometimes necessitating inconveniently distinct library preparation workflows within the same lab depending on the use case. Here we introduce a new PCR master mix that can be used for a wide array of input amounts, sample types, and workflows including whole genome sequencing (WGS), library prep for hybrid capture, and post-hybrid capture amplification. Using whole genome sequencing of different DNA sample types and species, we demonstrate high library yield, uniform genome coverage, and consistent library insert size across a broad range of cycle numbers for both picogram and high nanogram levels of DNA input. Next, we applied this PCR master mix to hybrid capture workflows, achieving sufficient library yield for capture and robust amplification of post-capture libraries in the presence of streptavidin beads. Using tissue-extracted DNA and cell-free DNA (cfDNA) samples, we demonstrated the critical importance of fidelity to enable highly precise single nucleotide variant (SNV) and indel calling. As part of a library prep workflow that maximizes conversion and introduces minimal errors, this PCR master mix can enable more accurate and cost-effective somatic variant detection. This new PCR master mix is designed to be readily applied to most DNA library prep applications with broad input amount and cycle number compatibility. Our data emphasizes the benefits of a high-fidelity PCR master mix as part of a high conversion library prep workflow for sensitive variant detection and high accuracy sequencing platforms.