Diff-seq: A high throughput sequencing-based mismatch detection assay for DNA variant enrichment and discovery.

ABSTRACT: Much of the within species genetic variation is in the form of single nucleotide polymorphisms (SNPs), typically detected by whole genome sequencing (WGS) or microarray-based technologies. However, WGS produces mostly uninformative reads that perfectly match the reference, while microarrays require genome-specific reagents. We have developed Diff-seq, a sequencing-based mismatch detection assay for SNP discovery without the requirement for specialized nucleic-acid reagents. Diff-seq leverages the Surveyor endonuclease to cleave mismatched DNA molecules that are generated after cross-annealing of a complex pool of DNA fragments. Sequencing libraries enriched for Surveyor-cleaved molecules result in increased coverage at the variant sites. Diff-seq detected all mismatches present in an initi

SUBMITTER: Aggeli D 

PROVIDER: S-EPMC5909455 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature