QUANTITATION OF 5-METHYLCYTOSINE BY ONE-DIMENSIONAL HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY

被引:13
作者
LEONARD, SA [1 ]
WONG, SC [1 ]
NYCE, JW [1 ]
机构
[1] E CAROLINA UNIV,SCH MED,DEPT MOLEC PHARMACOL & THERAPEUT,GREENVILLE,NC 27858
来源
JOURNAL OF CHROMATOGRAPHY | 1993年 / 645卷 / 01期
关键词
D O I
10.1016/0021-9673(93)80635-L
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A method for the quantitative analysis of DNA 5-methylcytosine by one-dimensional high-performance thin-layer chromatography using alkylamino modified silica (HPTLC-NH2) plates is described. 'rhe preparative method is simple, involving enzymatic digestion of DNA with micrococcal nuclease and phosphodiesterase II to 3'-monophosphate nucleosides, conversion by T4 polynucleotide kinase to P-32-labeled 3',5'-bisphosphate nucleosides, and chromatographic separation of nuclease P1-cleaved 5'-monophosphate nucleosides. The weak, basic anion exchanger property of the HPTLC-NH2 plate enables separation of multiple samples in one dimension, whereas traditional polyethyleneimine cellulose plates require development of individual samples in two dimensions for analysis of 5-methylcytosine.
引用
收藏
页码:189 / 192
页数:4
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