Rapid quantification of total genomic DNA methylation degree by short-end injection capillary zone electrophoresis |
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Authors: | Sotgia Salvatore Carru Ciriaco Franconi Flavia Fiori Paolo Battista Manca Salvatorica Pettinato Silvestro Magliona Stefania Ginanneschi Renzo Deiana Luca Zinellu Angelo |
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Institution: | Department of Biomedical Sciences, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43, Sassari, Italy. salvatore.sotgia@gmail.com |
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Abstract: | 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. |
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