title: Methylated CpG Island Amplification and Microarray (MCAM) for High-Throughput Analysis of DNA Methylation.
abstract: INTRODUCTIONThis protocol describes the use of methylated CpG island amplification (MCA) in combination with a microarray platform to analyze genome-wide DNA methylation in a high-throughput fashion. In this approach, termed MCAM, methylated CpG islands are selectively targeted using oligonucleotide adaptors after two rounds of digestion with a combination of methylation-sensitive and methylation-insensitive nucleases. They are then amplified using PCR. The resulting amplicons, representing the methylated fraction of the genome, are labeled with fluorochromes. Subsequently, a comparative hybridization of reference and test samples (typically normal and tumor DNA specimens) is done on a microarray platform.
authors.affiliation: Department of Leukemia, University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, USA.
authors.forename: Marcos R H
authors.initials: MR
authors.lastname: Estécio
authors.forename: Pearlly S
authors.initials: PS
authors.lastname: Yan
authors.forename: Tim H-M
authors.initials: TH
authors.lastname: Huang
authors.forename: Jean-Pierre J
authors.initials: JP
authors.lastname: Issa