Detection of low-frequency DNA variants by targeted sequencing of the Watson and Crick strands
Journal Title
Nature Biotechnology
Publication Type
e409
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 <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.
Publisher
NPG
WEHI Research Division(s)
Personalised Oncology
PubMed ID
33941929
Rights Notice
Refer to copyright notice on published article.


Creation Date: 2021-05-11 04:12:07
Last Modified: 2021-05-11 04:19:38
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