Quantitative detection of methylated SOCS-1 , a tumor suppressor gene, by a modified protocol of quantitative real time methylation-specific PCR using SYBR green and its use in early gastric cancer detection.
Although methylation-specific PCR (MSP) is a sensitive technique in the detection of DNA hypermethylation, it is not quantitative. Here we described a modified PCR protocol to quantify methylated SOCS-1 gene by real time MSP using SYBR green, which involves an additional PCR step after the 72 degrees C extension step. This modified protocol is also useful in the quantitative detection of methylated SOCS-1 gene in serum samples of gastric cancer patients.
Department of Medicine and Therapeutics, The Chinese University of Hong Kong, Shatin, NT, Hong Kong.
Michael W Y
MW
Chan
Eagle S H
ES
Chu
Ka-Fai
KF
To
Wai K
WK
Leung
D000328:Adult; D052160:Benzothiazoles; D014408:Biomarkers, Tumor / Q000097:blood*; D019175:DNA Methylation; D003959:Diamines; D005260:Female; D016147:Genes, Tumor Suppressor; D020022:Genetic Predisposition to Disease / Q000235:genetics; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000097:blood* / Q000378:metabolism*; D008297:Male; D008745:Methylation; D008875:Middle Aged; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D012097:Repressor Proteins / Q000097:blood* / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013274:Stomach Neoplasms / Q000097:blood* / Q000235:genetics / Q000378:metabolism*; D000071222:Suppressor of Cytokine Signaling 1 Protein; D050826:Suppressor of Cytokine Signaling Proteins
D052160:Benzothiazoles; D014408:Biomarkers, Tumor; D003959:Diamines; D047908:Intracellular Signaling Peptides and Proteins; D009930:Organic Chemicals; D011804:Quinolines; D012097:Repressor Proteins; C108944:SOCS1 protein, human; D000071222:Suppressor of Cytokine Signaling 1 Protein; D050826:Suppressor of Cytokine Signaling Proteins; C098022:SYBR Green I