Quantification of methylated DNA by HeavyMethyl duplex PCR.
The HeavyMethyl (HM) assay is a real-time PCR assay suitable for the qualitative and quantitative DNA methylation analysis of fresh, frozen, or formalin-fixed paraffin-embedded tissues and remote samples, such as serum, plasma, and urine. The HM uses a methylation-specific oligonucleotide blocker and a methylation-specific probe to achieve methylation-specific amplification and detection. A protocol for a duplex real-time PCR for the analysis of the methylation status of the GSTP1 exon1 in prostate tissue samples is presented. This chapter describes the preparation and analysis of a combined HM GSTP1 Exon1 and GSTP1 reference assay.
Distler
Jürgen
J
Epigenomics AG, Berlin, Germany.
D001483:Base Sequence; D018899:CpG Islands; D019175:DNA Methylation; D017931:DNA Primers / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D005091:Exons; D051549:Glutathione S-Transferase pi / Q000235:genetics; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D011471:Prostatic Neoplasms / Q000175:diagnosis / Q000201:enzymology / Q000235:genetics; D012015:Reference Standards
D017931:DNA Primers; D004273:DNA, Neoplasm; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi