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- Title
SMRT Sequencing for Parallel Analysis of Multiple Targets and Accurate SNP Phasing.
- Authors
Xiaoge Guo; Lehner, Kevin; O'Connell, Karen; Zhang, Jenny; Dave, Sandeep S.; Jinks-Robertson, Sue
- Abstract
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.
- Subjects
NUCLEOTIDE sequencing; GENETIC research; B cells
- Publication
G3: Genes | Genomes | Genetics, 2015, Vol 5, Issue 12, p2801
- ISSN
2160-1836
- Publication type
Article
- DOI
10.1534/g3.115.023317