High-throughput and cost-effective global DNA methylation assay by liquid chromatography-mass spectrometry.

ABSTRACT: An affordable and fast liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the accurate and precise determination of global DNA methylation levels in peripheral blood. Global DNA methylation extent was expressed as the ratio of methylated 2'-deoxycytidine (5MedC) to 2'-deoxyguanosine (dG), which were obtained after DNA extraction and hydrolysis and determined by positive electrospray LC-ESI-MS/MS. The cost-effective internal standards (15)N(3)-dC and (15)N(5)-dG were incorporated for the accurate quantification of 5MedC and dG, respectively. The desired nucleoside analytes were separated and eluted by LC within 2.5 min on a reverse phase column with a limit of detection of 1.4femtomole on column for 5MedC. Sample preparation in 96-well format has significantl

SUBMITTER: Li X 

PROVIDER: S-EPMC4439212 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature