Highly Sensitive Targeted Methylome Sequencing by Post-Bisulfite Adaptor Tagging

DNA Res. 2015 Feb;22(1):13-8. doi: 10.1093/dnares/dsu034. Epub 2014 Oct 16.

Abstract

The current gold standard method for methylome analysis is whole-genome bisulfite sequencing (WGBS), but its cost is substantial, especially for the purpose of multi-sample comparison of large methylomes. Shotgun bisulfite sequencing of target-enriched DNA, or targeted methylome sequencing (TMS), can be a flexible, cost-effective alternative to WGBS. However, the current TMS protocol requires a considerable amount of input DNA and hence is hardly applicable to samples of limited quantity. Here we report a method to overcome this limitation by using post-bisulfite adaptor tagging (PBAT), in which adaptor tagging is conducted after bisulfite treatment to circumvent bisulfite-induced loss of intact sequencing templates, thereby enabling TMS of a 100-fold smaller amount of input DNA with far fewer cycles of polymerase chain reaction than in the current protocol. We thus expect that the PBAT-mediated TMS will serve as an invaluable method in epigenomics.

Keywords: DNA methylation; massively parallel sequencing; target enrichment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • DNA Methylation / physiology*
  • Epigenesis, Genetic / physiology*
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Mice
  • Sulfites / chemistry

Substances

  • Sulfites
  • hydrogen sulfite