Improved tagmentation-based whole-genome bisulfite sequencing for input DNA from less than 100 mammalian cells

Epigenomics. 2015;7(1):47-56. doi: 10.2217/epi.14.76.

Abstract

Aim: To develop a whole-genome methylation sequencing method that fulfills the needs for studies using ultra-low-input DNA.

Materials & methods: The tagmentation-based whole-genome bisulfite sequencing (T-WGBS) technology is modified, enabling stable library construction with complexity from minimally 0.5 ng of initial genomic DNA, which equals less than 100 mammalian cells.

Results: We thoroughly assessed the performance of this T-WGBS method by sequencing the methylomes of a rice strain and pre-implantation embryos of rhesus monkey and compare to traditional WGBS approach, thereby demonstrating the efficacy of this new approach.

Conclusion: This new approach is highly attractive for the complete methylome analysis of very few cells, for example, mammalian pre-implantation embryos, or tiny human biopsy specimens.

Keywords: DNA methylation; T-WGBS; Tn5 transposase; WGBS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Methylation*
  • Genomics / methods
  • Macaca mulatta
  • Oryza / genetics
  • Sequence Analysis, DNA / methods*
  • Sulfites

Substances

  • Sulfites
  • hydrogen sulfite