Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands Export

@article{citeulike:4369810, abstract = {Precise mapping of DNA methylation patterns in CpG islands has become essential for understanding diverse biological processes such as the regulation of imprinted genes, X chromosome inactivation, and tumor suppressor gene silencing in human cancer. We describe a new method, MSP (methylation-specific PCR), which can rapidly assess the methylation status of virtually any group of CpG sites within a CpG island, independent of the use of methylation-sensitive restriction enzymes. This assay entails initial modification of DNA by sodium bisulfite, converting all unmethylated, but not methylated, cytosines to uracil, and subsequent amplification with primers specific for methylated versus unmethylated DNA. MSP requires only small quantities of DNA, is sensitive to 0.1\% methylated alleles of a given CpG island locus, and can be performed on DNA extracted from paraffin-embedded samples. MSP eliminates the false positive results inherent to previous PCR-based approaches which relied on differential restriction enzyme cleavage to distinguish methylated from unmethylated DNA. In this study, we demonstrate the use of MSP to identify promoter region hypermethylation changes associated with transcriptional inactivation in four important tumor suppressor genes (p16, p15, E-cadherin, and von Hippel-Lindau) in human cancer.}, author = {Herman, J. G. and Graff, J. R. and My\~{A}{\P}h\~{A}¤nen, S. and Nelkin, B. D. and Baylin, S. B.}, citeulike-article-id = {4369810}, citeulike-linkout-0 = {http://www.pnas.org/content/93/18/9821.abstract}, citeulike-linkout-1 = {http://www.pnas.org/content/93/18/9821.full.pdf}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/8790415}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=8790415}, day = {3}, issn = {0027-8424}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {chf, msp}, month = {September}, number = {18}, pages = {9821--9826}, posted-at = {2009-11-06 18:41:06}, priority = {0}, title = {Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands}, url = {http://www.pnas.org/content/93/18/9821.abstract}, volume = {93}, year = {1996} }

TY - JOUR ID - citeulike:4369810 L3 - citeulike-article-id:4369810 N2 - Precise mapping of DNA methylation patterns in CpG islands has become essential for understanding diverse biological processes such as the regulation of imprinted genes, X chromosome inactivation, and tumor suppressor gene silencing in human cancer. We describe a new method, MSP (methylation-specific PCR), which can rapidly assess the methylation status of virtually any group of CpG sites within a CpG island, independent of the use of methylation-sensitive restriction enzymes. This assay entails initial modification of DNA by sodium bisulfite, converting all unmethylated, but not methylated, cytosines to uracil, and subsequent amplification with primers specific for methylated versus unmethylated DNA. MSP requires only small quantities of DNA, is sensitive to 0.1% methylated alleles of a given CpG island locus, and can be performed on DNA extracted from paraffin-embedded samples. MSP eliminates the false positive results inherent to previous PCR-based approaches which relied on differential restriction enzyme cleavage to distinguish methylated from unmethylated DNA. In this study, we demonstrate the use of MSP to identify promoter region hypermethylation changes associated with transcriptional inactivation in four important tumor suppressor genes (p16, p15, E-cadherin, and von Hippel-Lindau) in human cancer. IS - 18 JF - Proceedings of the National Academy of Sciences of the United States of America SN - 0027-8424 EP - 9826 TI - Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands VL - 93 SP - 9821 KW - chf KW - msp AU - Herman, JG AU - Graff, JR AU - Myöhänen, S AU - Nelkin, BD AU - Baylin, SB PY - 1996/09/03/ UR - http://www.pnas.org/content/93/18/9821.abstract ER -