Authors
Daniel J. Weisenberger, Mihaela Campan, Tiffany I. Long, and Peter W. Laird
Summary
MethyLight is a sensitive, quantitative, TaqMan-based, real-time PCR assay for measuring DNA methylation profiles. The MethyLight-based data is presented as a percentage relative to an M.SssI-treated methylated DNA reference sample (PMR). We have extended the applicability of MethyLight technology to assay for the presence of the CpG island methylator phenotype (CIMP) in human colorectal cancer. We provide here a detailed, step-by-step protocol of the MethyLight assay for detection of CIMP with high sensitivity and specificity in colorectal cancer using a five marker panel composed of CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. Our criteria state that a sample with ≥3 of five markers posititve for methylation (PMR>10) is CIMP positive (CIMP+), while a sample with ≤2 of the five markers positive for methylation is considered CIMP negative (CIMP-).Further details
The protocol was published on Protocol Exchange in 2006. To see the entire protocol, click on the source link.Advertisement
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