Novel procedure for the detection of 5-methylcytosine.
Bisulfite converts non-methylated cytosine in DNA to uracil leaving 5-methylcytosine unaltered. In this communication, we present a new approach omitting the conventional PCR amplification step. Bisulfite-converted methylated DNA is directly sequenced. The effectiveness of the new protocol is demonstrated by using it for the detection of 5-methylation of cytosine residues introduced by three different DNA methyltransferases (M.HaeIII, M.HpaII and M.HhaI). A simple experimental system useful to determine the sequence specificity of DNA methyltransferases is also presented.
Radlińska
M
M
Department of Virology, Warsaw University, Poland.
Skowronek
K
K
D044503:5-Methylcytosine; D001482:Base Composition; D003596:Cytosine / Q000031:analogs & derivatives* / Q000032:analysis; D004247:DNA / Q000737:chemistry; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000378:metabolism; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D017422:Sequence Analysis, DNA; D013447:Sulfites / Q000378:metabolism
D013447:Sulfites; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA; D015254:DNA Modification Methylases; C042345:hydrogen sulfite