SMRT Sequencing for Parallel Analysis of Multiple Targets and Accurate SNP Phasing.

Published online

Journal Article

Single-molecule real-time (SMRT) sequencing generates much longer reads than other widely used next-generation (next-gen) sequencing methods, but its application to whole genome/exome analysis has been limited. Here, we describe the use of SMRT sequencing coupled with barcoding to simultaneously analyze one or a small number of genomic targets derived from multiple sources. In the budding yeast system, SMRT sequencing was used to analyze strand-exchange intermediates generated during mitotic recombination and to analyze genetic changes in a forward mutation assay. The general barcoding-SMRT approach was then extended to diffuse large B-cell lymphoma primary tumors and cell lines, where detected changes agreed with prior Illumina exome sequencing. A distinct advantage afforded by SMRT sequencing over other next-gen methods is that it immediately provides the linkage relationships between SNPs in the target segment sequenced. The strength of our approach for mutation/recombination studies (as well as linkage identification) derives from its inherent computational simplicity coupled with a lack of reliance on sophisticated statistical analyses.

Full Text

Duke Authors

Cited Authors

  • Guo, X; Lehner, K; O'Connell, K; Zhang, J; Dave, SS; Jinks-Robertson, S

Published Date

  • October 23, 2015

Published In

Volume / Issue

  • 5 / 12

Start / End Page

  • 2801 - 2808

PubMed ID

  • 26497143

Pubmed Central ID

  • 26497143

Electronic International Standard Serial Number (EISSN)

  • 2160-1836

Digital Object Identifier (DOI)

  • 10.1534/g3.115.023317

Language

  • eng

Conference Location

  • United States