A bisulfite method of 5-methylcytosine mapping that minimizes template degradation.
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.
Raizis
A M
AM
Friedrich Miescher Institut, Basel, Switzerland.
Schmitt
F
F
Jost
J P
JP
D044503:5-Methylcytosine; D001483:Base Sequence; D003596:Cytosine / Q000031:analogs & derivatives* / Q000032:analysis; D004247:DNA / Q000737:chemistry*; D015254:DNA Modification Methylases / Q000378:metabolism; D017931:DNA Primers; D006863:Hydrogen-Ion Concentration; D008745:Methylation; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013447:Sulfites / Q000494:pharmacology*; D013698:Templates, Genetic
D017931:DNA Primers; D013447:Sulfites; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA; D015254:DNA Modification Methylases; C042345:hydrogen sulfite