DNA ELECTROPHORESIS IN UNCROSS-LINKED POLYACRYLAMIDE SOLUTION, STUDIED BY EPIFLUORESCENCE MICROSCOPY

被引:19
作者
RAMPINO, NJ
CHRAMBACH, A
机构
[1] Section on Macromolecular Analysis, Laboratory of Theoretical and Physical Biology, National Institute of Child Health and Human Development, Bethesda
来源
JOURNAL OF CHROMATOGRAPHY | 1992年 / 596卷 / 01期
关键词
D O I
10.1016/0021-9673(92)80217-I
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Electrophoresis of human DNA fragments (approximately 1 . 10(5) to 1 . 10- bases in size) was conducted in a solution of uncross-linked polyacrylamide contained in a horizontally mounted 1 mm diameter glass tube and monitored by epifluorescence microscopy. In presence of the polymer, molecular conformations described as a "trailing network" of DNA and a globular "head" were observed. The migration velocity varies between species differing in the size of the "head", and in the ratio between the size of the "head" and that of the trailing "network". By contrast, in pure buffer, lambda-phage DNA migrates in a globular form at a mobility consistent with known macroscopic data. When electrophoresis in the polymer solution of an agarose plug preparation of Schizosaccharomyces pombe DNA was carried out after melting at 70-degrees-C, a migrating DNA-agarose complex was observed. The complex was not fully dissociated by an agarose-hydrolyzing enzyme (Gelase).
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页码:141 / 149
页数:9
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