An optimized targeted Next-Generation Sequencing approach for sensitive detection of single nucleotide variants.

•NGS based detection of low-level SNVs is feasible with sensitivities up to 10−4.•PCR-induced bias could be significantly reduced by the choice of adequate enzymes.•The prevalent transition vs. transversion bias affects site-specific detection limits.•Results from clinical data validated the feasibility of NGS-based MRD detection.•Results help to select suitable biomarkers for MRD quantification.