title: Determination of global DNA methylation level by capillary electrophoresis using octyl-modified quaternized cellulose as an electrolyte additive.
abstract: A new cellulose derivative, octyl-modified quaternized cellulose (OMQC), was synthesized and used as electrolyte additive for the analysis of 5-methylcytosine by capillary electrophoresis with UV detection. While added in the background electrolyte, OMQC carrying octyl groups and quaternary ammonium groups exhibited dynamic coating ability. Capillary coated with OMQC was able to generate a stable anodal electro-osmotic flow even at pH 12.0. After several running conditions were optimized, a new method for quantification of genomic methylation level was developed on the basis of hydrolysis of DNA by formic acid and separation of nucleic acid bases by capillary electrophoresis. Cytosine and 5-methylcytosine were separated with a resolution near 4.0 in less than 10 min. The detection limits (S/N = 3) were 1.1 and 1.5 μg/mL for cytosine and 5-methylcytosine, respectively.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, China.
authors.forename: Dihui
authors.initials: D
authors.lastname: Huang
authors.forename: Qin
authors.initials: Q
authors.lastname: Yang
authors.forename: Shanxia
authors.initials: S
authors.lastname: Jin
authors.forename: Qianchun
authors.initials: Q
authors.lastname: Deng
authors.forename: Ping
authors.initials: P
authors.lastname: Zhou
mesh_terms: D044503:5-Methylcytosine / Q000737:chemistry; D002482:Cellulose / Q000737:chemistry*; D003596:Cytosine / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D019175:DNA Methylation; D004573:Electrolytes / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D006868:Hydrolysis
chemical_list: D004573:Electrolytes; D044503:5-Methylcytosine; D003596:Cytosine; D002482:Cellulose; D004247:DNA