Ultra-low-input, tagmentation-based whole-genome bisulfite sequencing

Genome Res. 2012 Jun;22(6):1139-43. doi: 10.1101/gr.136242.111. Epub 2012 Mar 30.

Abstract

We have adapted transposase-based in vitro shotgun library construction ("tagmentation") for whole-genome bisulfite sequencing. This method, Tn5mC-seq, enables a >100-fold reduction in starting material relative to conventional protocols, such that we generate highly complex bisulfite sequencing libraries from as little as 10 ng of input DNA, and ample useful sequences from 1 ng of input DNA. We demonstrate Tn5mC-seq by sequencing the methylome of a human lymphoblastoid cell line to ∼8.6× high-quality coverage of each strand.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Sequence
  • Cell Line, Transformed
  • CpG Islands
  • Cytosine / chemistry
  • DNA / drug effects
  • DNA Methylation
  • Gene Library*
  • Genome, Human*
  • Humans
  • Lymphocytes / pathology
  • Lymphocytes / physiology
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA / methods*
  • Sulfites / pharmacology
  • Transposases / genetics

Substances

  • Sulfites
  • Cytosine
  • DNA
  • Transposases
  • hydrogen sulfite