Genome-scale validation of deep-sequencing libraries

PLoS One. 2008;3(11):e3713. doi: 10.1371/journal.pone.0003713. Epub 2008 Nov 12.

Abstract

Chromatin immunoprecipitation followed by high-throughput (HTP) sequencing (ChIP-seq) is a powerful tool to establish protein-DNA interactions genome-wide. The primary limitation of its broad application at present is the often-limited access to sequencers. Here we report a protocol, Mab-seq, that generates genome-scale quality evaluations for nucleic acid libraries intended for deep-sequencing. We show how commercially available genomic microarrays can be used to maximize the efficiency of library creation and quickly generate reliable preliminary data on a chromosomal scale in advance of deep sequencing. We also exploit this technique to compare enriched regions identified using microarrays with those identified by sequencing, demonstrating that they agree on a core set of clearly identified enriched regions, while characterizing the additional enriched regions identifiable using HTP sequencing.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Chromatin Immunoprecipitation
  • Genome
  • Genomic Library*
  • Genomics / methods*
  • Humans
  • Models, Biological
  • Oligonucleotide Array Sequence Analysis