Benjamin P. Song, Surbhi Jain, Selena Y. Lin, Quan Chen, Timothy M

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Detection of Hypermethylated Vimentin in Urine of Patients with Colorectal Cancer Benjamin P. Song, Surbhi Jain, Selena Y. Lin, Quan Chen, Timothy M. Block, Wei Song, Dean E. Brenner, Ying-Hsiu Su The Journal of Molecular Diagnostics Volume 14, Issue 2, Pages 112-119 (March 2012) DOI: 10.1016/j.jmoldx.2011.12.003 Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Development of a two-step MethyLight PCR assay for the mVIM gene. A: Schematic location of the CpG sites generated by the Methyl Primer Express software in the promoter and the first exon regions of the vimentin gene (GenBank AL133415) and the locations of the forward and reverse primer regions of the MSP29 methylation-specific PCR assay37 are indicated. Each vertical line represents a CpG site. B: Schematic diagrams of the two-step nested quantitative MethyLight assay. The green line represents the target sequence. For the two-step PCR assay, the forward and reverse primers of the first PCR cycle include the targeted sequence and an artificial sequence (orange). The second PCR cycle has a forward and a reverse primer (with mostly artificial sequences shown in orange) and a TaqMan probe. C: Amplification curves of the mVIM MethyLight assay VIM29R_LNA. The amplification curves and the linear standard were generated from a reconstituted standard as described in Materials and Methods. The numbers of copies of positive control DNA, negative control DNA (Neg), and H2O are indicated. The Journal of Molecular Diagnostics 2012 14, 112-119DOI: (10.1016/j.jmoldx.2011.12.003) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions