Detection of low-frequency DNA variants by targeted sequencing of the Watson and Crick strands
- 3 May 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Biotechnology
- Vol. 39 (10), 1220-1227
- https://doi.org/10.1038/s41587-021-00900-z
Abstract
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 100-fold.Keywords
Funding Information
- U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (T32-GM007309, T32-GM007309)
- Victorian Cancer Agency (CRF14007, CRF14007)
- John Templeton Foundation
- Conrad N. Hilton Foundation
- Virginia and D.K. Ludwig Fund for Cancer Research
- U.S. Department of Health & Human Services | NIH | National Cancer Institute (U01 CA152753, P50CA228991, U01CA200469, R37 CA230400-01, CA62924, CA210170)
- Lustgarten Foundation
- Billi and Bernie Marcus Foundation
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