Open AccessNew library construction method for single-cell genomes
Author(s) -
Xi Liu,
A. N. Belyaev,
Sandra L. Spurgeon,
Xiaohui Wang,
Haibiao Gong,
Robert Aboukhalil,
Richard A. Fekete
Publication year - 2017
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0181163
Subject(s) - genome , genomic library , computational biology , dna sequencing , transposition (logic) , biology , genomics , computer science , genetics , dna , base sequence , gene , artificial intelligence
A central challenge in sequencing single-cell genomes is the accurate determination of point mutations, phasing of these mutations, and identifying copy number variations with few assumptions. Ideally, this is accomplished under as low sequencing coverage as possible. Here we report our attempt to meet these goals with a novel library construction and library amplification methodology. In our approach, single-cell genomic DNA is first fragmented with saturated transposition to make a primary library that uniformly covers the whole genome by short fragments. The library is then amplified by a carefully optimized PCR protocol in a uniform and synchronized fashion for next-generation sequencing. Each step of the protocol can be quantitatively characterized. Our shallow sequencing data show that the library is tightly distributed and is useful for the determination of copy number variations.