Detection of low-frequency DNA variants by targeted sequencing of the Watson and Crick strands
- Cohen, JD; Douville, C; Dudley, JC; Mog, BJ; Popoli, M; Ptak, J; Dobbyn, L; Silliman, N; Schaefer, J; Tie, J; Gibbs, P; Tomasetti, C; Papadopoulos, N; Kinzler, KW; Vogelstein, B;
- Journal Title
- Nature Biotechnology
- Publication Type
- Identification and quantification of low-frequency mutations remain challenging despite improvements in the baseline error rate of next-generation sequencing technologies. Here, we describe a method, termed SaferSeqS, that addresses these challenges by (1) efficiently introducing identical molecular barcodes in the Watson and Crick strands of template molecules and (2) enriching target sequences with strand-specific PCR. The method achieves high sensitivity and specificity and detects variants at frequencies below 1 in 100,000 DNA template molecules with a background mutation rate of <5 × 10(-7) mutants per base pair (bp). We demonstrate that it can evaluate mutations in a single amplicon or simultaneously in multiple amplicons, assess limited quantities of cell-free DNA with high recovery of both strands and reduce the error rate of existing PCR-based molecular barcoding approaches by >100-fold.
- WEHI Research Division(s)
- Personalised Oncology
- PubMed ID
- Publisher's Version
- Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2021-05-11 04:12:07Last Modified: 2021-05-11 04:19:38