TY - JOUR
T1 - Evaluation of Global Genomic DNA Methylation in Human Whole Blood by Capillary Electrophoresis UV Detection
AU - Zinellu, Angelo
AU - Sotgiu, Elisabetta
AU - Assaretti, Stefano
AU - Sotgia, Salvatore
AU - Paliogiannis, Panagiotis
AU - Pintus, Gianfranco
AU - Mangoni, Arduino
AU - Carru, Ciriaco
PY - 2017
Y1 - 2017
N2 - Alterations in global DNA methylation are implicated in various pathophysiological processes. )e development of simple and quick, yet robust, methods to assess DNA methylation is required to facilitate its measurement and interpretation in clinical practice. We describe a highly sensitive and reproducible capillary electrophoresis method with UV detection for the separation and detection of cytosine and methylcytosine, after formic acid hydrolysis of DNA extracted from human whole blood. Hydrolysed samples were dried and resuspended with water and directly injected into the capillary without sample derivatization procedures. )e use of a run bu8er containing 50 mmol/L BIS-TRIS propane (BTP) phosphate bu8er at pH 3.25 and 60 mmol/L sodium acetate bu8er at pH 3.60 (4 : 1, v/v) allowed full analyte identiCcation within 11 min. Precision tests indicated an elevated reproducibility with an interassay CV of 1.98% when starting from 2 μg of the extracted DNA. )e method was successfully tested by measuring the DNA methylation degree both in healthy volunteers and in reference calf thymus DNA.
AB - Alterations in global DNA methylation are implicated in various pathophysiological processes. )e development of simple and quick, yet robust, methods to assess DNA methylation is required to facilitate its measurement and interpretation in clinical practice. We describe a highly sensitive and reproducible capillary electrophoresis method with UV detection for the separation and detection of cytosine and methylcytosine, after formic acid hydrolysis of DNA extracted from human whole blood. Hydrolysed samples were dried and resuspended with water and directly injected into the capillary without sample derivatization procedures. )e use of a run bu8er containing 50 mmol/L BIS-TRIS propane (BTP) phosphate bu8er at pH 3.25 and 60 mmol/L sodium acetate bu8er at pH 3.60 (4 : 1, v/v) allowed full analyte identiCcation within 11 min. Precision tests indicated an elevated reproducibility with an interassay CV of 1.98% when starting from 2 μg of the extracted DNA. )e method was successfully tested by measuring the DNA methylation degree both in healthy volunteers and in reference calf thymus DNA.
UR - http://www.scopus.com/inward/record.url?scp=85039449954&partnerID=8YFLogxK
U2 - 10.1155/2017/4065892
DO - 10.1155/2017/4065892
M3 - Article
SN - 2090-8873
VL - 2017
JO - Journal of Analytical Methods in Chemistry
JF - Journal of Analytical Methods in Chemistry
M1 - 4065892
ER -