Rapid quantification of total genomic DNA methylation degree by short-end injection capillary zone electrophoresis.
We propose a new capillary zone electrophoresis method applying short-end injection technique for the fast evaluation of methylcystosine/total cytosine ratio after acidic DNA hydrolysis. By short-end injection and by using a 100 mmol/l Tris solution titrated with 1 mol/l phosphoric acid to pH 3.75 as background electrolyte, cytosine and methylcytosine were separated with a good resolution in less than 1.5 min. Stepwise multiple linear regression with DNA methylation degree as the dependent variable and age, cysteine, homocysteine and methionine as independent variables, showed a negative association with age and that total cysteine is the most important determinant of DNA methylation.
Sotgia
Salvatore
S
Department of Biomedical Sciences, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43, Sassari, Italy. salvatore.sotgia@gmail.com
Carru
Ciriaco
C
Franconi
Flavia
F
Fiori
Paolo Battista
PB
Manca
Salvatorica
S
Pettinato
Silvestro
S
Magliona
Stefania
S
Ginanneschi
Renzo
R
Deiana
Luca
L
Zinellu
Angelo
A
D004247:DNA / Q000378:metabolism*; D019175:DNA Methylation; D018744:DNA, Plant; D019075:Electrophoresis, Capillary / Q000379:methods*; D016678:Genome / Q000502:physiology*; D006868:Hydrolysis; D013696:Temperature
D018744:DNA, Plant; D004247:DNA