Validation of MSRE-PCR results with selected genes

Copyright information:Taken from "MSRE-PCR for analysis of gene-specific DNA methylation"Nucleic Acids Research 2005;33(10):e93-e93.Published online 8 Jun 2005PMCID:PMC1145194.© The Author 2005. Published by Oxford University Press. All rights reserved () Methylation-specific PCR (MSP) assay of four genes in MCF-7 cells (duplicate reactions for each gene). Primers for unmethylated (U) and methylated (M) DNA of corresponding CpG islands were used. Fragments amplified for MSP are located within regions analyzed by MSRE-PCR. () Bisulfite sequencing of a calcitonin promoter fragment located within a larger fragment used for MSRE-PCR. Probability of methylation for each of 19 CpG dinucleotides for MCF-7 (open squares) and T47D (filled squares) is plotted. Vertical arrows mark the position of CpG dinucleotides; arrowheads indicate CpG dinucleotides within the Hin6I recognition site. Horizontal arrows mark the position of PCR primers for MSRE-PCR (dashed arrows, primers A and B) and primers for amplification of bisulfite-modified DNA (dotted arrows, primers a and b). () Northern blot analysis of gene expression in MCF-7, T47D and MDA-MB-231 cells. β-Actin was used as a loading control.