Post-conversion targeted capture of modified cytosines in mammalian and plant genomes

Nucleic Acids Res. 2015 Jul 13;43(12):e81. doi: 10.1093/nar/gkv244. Epub 2015 Mar 26.

Abstract

We present a capture-based approach for bisulfite-converted DNA that allows interrogation of pre-defined genomic locations, allowing quantitative and qualitative assessments of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) at CG dinucleotides and in non-CG contexts (CHG, CHH) in mammalian and plant genomes. We show the technique works robustly and reproducibly using as little as 500 ng of starting DNA, with results correlating well with whole genome bisulfite sequencing data, and demonstrate that human DNA can be tested in samples contaminated with microbial DNA. This targeting approach will allow cell type-specific designs to maximize the value of 5mC and 5hmC sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5-Methylcytosine / analysis*
  • Alleles
  • Animals
  • Cell Line
  • Cytosine / analogs & derivatives*
  • Cytosine / analysis
  • DNA Methylation
  • Genome, Plant*
  • Genomics / methods
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Mice
  • Polymorphism, Single Nucleotide
  • Sequence Analysis, DNA / methods*
  • Sulfites

Substances

  • Sulfites
  • 5-hydroxymethylcytosine
  • 5-Methylcytosine
  • Cytosine
  • hydrogen sulfite