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BMC Genomics. 2017 Oct 23;18(1):811. doi: 10.1186/s12864-017-4159-7.

dCATCH-Seq: improved sequencing of large continuous genomic targets with double-hybridization.

Author information

1
HudsonAlpha Institute for Biotechnology, Huntsville, USA.
2
HudsonAlpha Institute for Biotechnology, Huntsville, USA. dabsher@hudsonalpha.org.
3
Ubiquity Genomics Inc., Huntsville, USA. dabsher@hudsonalpha.org.

Abstract

BACKGROUND:

Targeted sequencing is a powerful tool with broad application in both basic and translational sciences. Relatively low on-target rates for most current targeted sequencing studies influence the required coverage and data quality for subsequent applications.

RESULTS:

We present an improved targeted sequencing method that uses two rounds of in solution hybridization with probes synthesized from genomic clone templates, termed dCATCH-Seq. Independent captures of two large continuous genomic regions across three cell types within the human major histocompatibility complex (MHC) that spans ~3.5 Mb and a ~250 kb region on chromosome 11 demonstrated that dCATCH-Seq was highly reproducible with ~95% capture specificity. Comprehensive analyses of sequencing data generated using the dCATCH-Seq approach also showed high accuracy in the detection of genetic variants and HLA typing. The double hybridization capture approach can also be coupled with bisulfite sequencing for DNA methylation profiling of both CpG and non-CpG sites.

CONCLUSIONS:

Altogether, dCATCH-Seq is a powerful and scalable targeted sequencing approach to investigate both genetic and epigenetic features.

KEYWORDS:

Bac; DNA methylation; Hla; Hybridization; Mhc; Targeted sequencing

PMID:
29061109
PMCID:
PMC5653981
DOI:
10.1186/s12864-017-4159-7
[Indexed for MEDLINE]
Free PMC Article

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