Multiplexed Illumina sequencing libraries from picogram quantities of DNA

BMC Genomics

Methodology Article

Michael Tolstorukov¹  Sarah K Bowman²  Aimee M Deaton²  Robert E Kingston²  Matthew D Simon³  Mark L Borowsky⁴ 

[1] Center for Biomedical Informatics, Harvard Medical School, 02115, Boston, MA, USA;Division of Genetics, Brigham and Women’s Hospital, 02115, Boston, MA, USA;Department of Molecular Biology, Massachusetts General Hospital, 02114, Boston, MA, USA;Department of Molecular Biology, Massachusetts General Hospital, and Department of Genetics, Harvard Medical School, 02114, Boston, MA, USA;Department of Molecular Biology, Massachusetts General Hospital, and Department of Genetics, Harvard Medical School, 02114, Boston, MA, USA;Department of Molecular Biophysics and Biochemistry and Chemical Biology Institute, Yale University, 06516, West Haven, CT, USA;Department of Molecular Biology, Massachusetts General Hospital, and Department of Genetics, Harvard Medical School, 02114, Boston, MA, USA;Novartis Institutes for BioMedical Research, 02139, Cambridge, MA, USA;

关键词: Illumina;    ChIP-seq;    Multiplex;    Barcoding;    Library preparation;   

DOI  :  10.1186/1471-2164-14-466

 received in 2012-09-26, accepted in 2013-07-06,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundHigh 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.ResultsWe 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.ConclusionsApplication of this method allows whole genome studies from samples where material or yields are limiting.

【 授权许可】

CC BY   
© Bowman et al.; licensee BioMed Central Ltd. 2013