Quantitative methylation analysis using methylation-sensitive single-nucleotide primer extension (Ms-SNuPE).
Methylation-sensitive single-nucleotide primer extension (Ms-SNuPE) is a technique that allows for rapid and simultaneous quantitation of the degree of methylation at several CpG sites. Treatment of genomic DNA with sodium bisulfite is used to convert unmethylated cytosine to uracil while leaving 5-methylcytosine unchanged. A strand-specific polymerase chain reaction product is then generated to provide a suitable DNA template for quantitative methylation analysis using Ms-SNuPE. Single-nucleotide primer extension is performed with oligonucleotide(s) designed to hybridize immediately upstream of the CpG site(s) being analyzed. The Ms-SNuPE technique can be adapted for high-throughput methylation analysis and therefore represents a novel approach for rapid quantitation of cytosine methylation suitable for a wide range of biological investigations.
Gonzalgo
Mark L
ML
Department of Biochemistry and Molecular Biology, Urologic Cancer Research Laboratory, USC/Norris Comprehensive Cancer Center, Keck School of Medicine, Los Angeles, CA 90089-9181, USA.
Jones
Peter A
PA
D018899:CpG Islands; D004247:DNA / Q000302:isolation & purification; D019175:DNA Methylation; D017931:DNA Primers; D015342:DNA Probes; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D013447:Sulfites / Q000378:metabolism; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D015342:DNA Probes; D013447:Sulfites; D004247:DNA