Verification of complete bisulfite modification using Calponin-specific primer sets.
OBJECTIVE
To verify complete bisulfite modification.


METHODS
Bisulfite modified DNA was screened by PCR with Calponin-specific primer sets prior to detecting methylation status of 14-3-3sigma by methylation-specific PCR.


RESULTS
False positive methylation was obtained from incomplete bisulfite modification. The lower detection limit of wild type Calponin-specific primer is approximately 0.375 ng of unmodified DNA.


CONCLUSIONS
Verification of complete bisulfite modification by Calponin-specific primer sets can reduce misinterpretation of methylation.
Sriraksa
Ruethairat
R
Graduate School, Khon Kaen University, Khon Kaen 40002, Thailand.
Chaopatchayakul
Patimaporn
P
Jearanaikoon
Patcharee
P
Leelayuwat
Chanvit
C
Limpaiboon
Temduang
T
D002135:Calcium-Binding Proteins / Q000235:genetics*; D019175:DNA Methylation; D017931:DNA Primers / Q000378:metabolism*; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013447:Sulfites / Q000378:metabolism*; D000096985:Calponins
D002135:Calcium-Binding Proteins; D017931:DNA Primers; D008840:Microfilament Proteins; D013447:Sulfites; D000096985:Calponins; C042345:hydrogen sulfite