Quantitative gene analysis of methylation and expression (Q-GAME) in fresh or fixed cells and tissues.

ABSTRACT: Epigenetic regulation of gene expression by DNA methylation is a central mechanism governing the silencing of tumor suppressor genes in many forms of cancer. Current methods have not proven optimal for the quantitative analysis of DNA methylation and corresponding in situ protein expression within cells in small specimens like skin biopsies. We have overcome this limitation by combining and modifying several techniques: target cell enrichment, DNA micro-isolation, one-step denaturation/bisulphite conversion/in-column desulphonation, specially designed PCR amplification, pyrosequencing and multispectral image analysis. Using this approach optimized for small samples, we can quantify minor alterations in gene methylation and protein expression using minimal amounts of tissue. Comparative stu

SUBMITTER: Wu J 

PROVIDER: S-EPMC4331060 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature