OBJECTIVES: To verify complete bisulfite modification. DESIGN AND 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. Copyright 2009 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
OBJECTIVES: To verify complete bisulfite modification. DESIGN AND 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. Copyright 2009 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
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