Highly sensitive targeted methylome sequencing by post-bisulfite adaptor tagging

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25 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)13-18
Number of pages6
JournalDNA Research
Volume22
Issue number1
DOIs
Publication statusPublished - 2015

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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