A bisulfite method of 5-methylcytosine mapping that minimizes template degradation

Anal Biochem. 1995 Mar 20;226(1):161-6. doi: 10.1006/abio.1995.1204.


The bisulfite method is a highly sensitive approach to 5-methylcytosine mapping that utilizes the capability of the polymerase chain reaction to exponentially amplify DNA. We have observed that the bisulfite reaction results in a significant level of template degradation due to DNA depurination. Furthermore, our data suggest that the DNA fragmentation which occurs limits the sensitivity of the method. We describe a simple solution to limit degradation of the DNA template.

MeSH terms

  • 5-Methylcytosine
  • Base Sequence
  • Cytosine / analogs & derivatives*
  • Cytosine / analysis
  • DNA / chemistry*
  • DNA Modification Methylases / metabolism
  • DNA Primers
  • Hydrogen-Ion Concentration
  • Methylation
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sulfites / pharmacology*
  • Templates, Genetic


  • DNA Primers
  • Sulfites
  • 5-Methylcytosine
  • Cytosine
  • DNA
  • DNA Modification Methylases
  • hydrogen sulfite