Multiplexed Illumina sequencing libraries from picogram quantities of DNA

被引：60

作者：

Bowman, Sarah K. ^{[1
,2
]}

Simon, Matthew D. ^{[1
,2
]}

Deaton, Aimee M. ^{[1
,2
]}

Tolstorukov, Michael ^{[3
,4
]}

Borowsky, Mark L. ^{[1
,2
]}

Kingston, Robert E. ^{[1
,2
]}

机构：

[1] Massachusetts Gen Hosp, Dept Mol Biol, Boston, MA 02114 USA

[2] Harvard Univ, Sch Med, Dept Genet, Boston, MA 02114 USA

[3] Harvard Univ, Sch Med, Ctr Biomed Informat, Boston, MA 02115 USA

[4] Brigham & Womens Hosp, Div Genet, Boston, MA 02115 USA

来源：

BMC GENOMICS | 2013年 / 14卷

关键词：

Illumina; ChIP-seq; Multiplex; Barcoding; Library preparation; CHIP-SEQ; GENOME; AMPLIFICATION;

D O I：

10.1186/1471-2164-14-466

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Background: High throughput sequencing is frequently used to discover the location of regulatory interactions on chromatin. However, techniques that enrich DNA where regulatory activity takes place, such as chromatin immunoprecipitation (ChIP), often yield less DNA than optimal for sequencing library preparation. Existing protocols for picogram-scale libraries require concomitant fragmentation of DNA, pre-amplification, or long overnight steps. Results: We report a simple and fast library construction method that produces libraries from sub-nanogram quantities of DNA. This protocol yields conventional libraries with barcodes suitable for multiplexed sample analysis on the Illumina platform. We demonstrate the utility of this method by constructing a ChIP-seq library from 100 pg of ChIP DNA that demonstrates equivalent genomic coverage of target regions to a library produced from a larger scale experiment. Conclusions: Application of this method allows whole genome studies from samples where material or yields are limiting.

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