Analysis of DNA methylation in FFPE tissues using the MethyLight technology.

Ashraf Dallol, Waleed Al-Ali, Amina Al-Shaibani, Fahd Al-Mulla

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Novel biomarkers are sought after by mining DNA extracted from formalin-fixed, paraffin-embedded (FFPE) tissues. Such tissues offer the great advantage of often having complete clinical data (including survival), as well as the tissues are amenable for laser microdissection targeting specific tissue areas. Downstream analysis of such DNA includes mutational screens and methylation profiling. Screening for mutations by sequencing requires a significant amount of DNA for PCR and cycle sequencing. This is self-inhibitory if the gene screened has a large number of exons. Profiling DNA methylation using the MethyLight technology circumvents this problem and allows for the mining of several biomarkers from DNA extracted from a single microscope slide of the tissue of interest. We describe in this chapter a detailed protocol for MethyLight and its use in the determination of CpG Island Methylator Phenotype status in FFPE colorectal cancer samples.

Original languageEnglish
Pages (from-to)191-204
Number of pages14
JournalMethods in molecular biology (Clifton, N.J.)
Volume724
DOIs
Publication statusPublished - 15 Jun 2011

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