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Multiplexed Illumina sequencing libraries from picogram quantities of DNA

Bowman, SK, Simon, MD, Deaton, AM, Tolstorukov, M, Borowsky, ML and Kingston, RE (2013) Multiplexed Illumina sequencing libraries from picogram quantities of DNA. BMC Genomics.


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. 2013 Bowman et al.; licensee BioMed Central Ltd

Item Type: Article
Additional Information: pubid: 153 nvp_institute: NIBR contributor_address: (Bowman, Simon, Deaton, Tolstorukov, Borowsky, Kingston) Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114, United States (Bowman, Simon, Deaton, Borowsky, Kingston) Department of Genetics, Harvard Medical School, Boston, MA 02114, United States (Tolstorukov) Center for Biomedical Informatics, Harvard Medical School, Boston, MA 02115, United States (Tolstorukov) Division of Genetics, Brigham and Women's Hospital, Boston, MA 02115, United States (Simon) Department of Molecular Biophysics and Biochemistry, Chemical Biology Institute, Yale University, West Haven, CT 06516, United States (Borowsky) Novartis Institutes for BioMedical Research, Cambridge, MA 02139, United States
Date Deposited: 13 Oct 2015 13:12
Last Modified: 13 Oct 2015 13:12