Exome sequencing from nanogram amounts of starting DNA: comparing three approaches

PLoS One. 2014 Jul 3;9(7):e101154. doi: 10.1371/journal.pone.0101154. eCollection 2014.


Hybridization-based target enrichment protocols require relatively large starting amounts of genomic DNA, which is not always available. Here, we tested three approaches to pre-capture library preparation starting from 10 ng of genomic DNA: (i and ii) whole-genome amplification of DNA samples with REPLI-g (Qiagen) and GenomePlex (Sigma) kits followed by standard library preparation, and (iii) library construction with a low input oriented ThruPLEX kit (Rubicon Genomics). Exome capture with Agilent SureSelectXT2 Human AllExon v4+UTRs capture probes, and HiSeq2000 sequencing were performed for test libraries along with the control library prepared from 1 µg of starting DNA. Tested protocols were characterized in terms of mapping efficiency, enrichment ratio, coverage of the target region, and reliability of SNP genotyping. REPLI-g- and ThruPLEX-FD-based protocols seem to be adequate solutions for exome sequencing of low input samples.

Publication types

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

MeSH terms

  • DNA / blood
  • DNA / genetics*
  • Exome*
  • Gene Library
  • Genomics / methods
  • Genotyping Techniques / methods
  • Humans
  • Sample Size
  • Sequence Analysis, DNA / methods*


  • DNA

Grant support

This work was supported by the 7th framework programme of the European Union (ADAMS) [242257, FP7-HEALTH-2009]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.