SEPARATION AND ANALYSIS OF DNA-SEQUENCE REACTION-PRODUCTS BY CAPILLARY GEL-ELECTROPHORESIS

被引:168
作者
COHEN, AS [1 ]
NAJARIAN, DR [1 ]
KARGER, BL [1 ]
机构
[1] NORTHEASTERN UNIV,BARNETT INST,BOSTON,MA 02115
来源
JOURNAL OF CHROMATOGRAPHY | 1990年 / 516卷 / 01期
关键词
D O I
10.1016/S0021-9673(01)90203-1
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
This paper demonstrates the potential of capillary gel electrophoresis with laser induced fluorescence detection as a tool for DNA sequence determination. Both synthetic oligonucleotides and single-stranded phage DNA were utilized as templates in the standard chain termination procedure. Primer molecules were tagged at the 5' end with the fluorescent dye, JOE. First, baseline resolution of a dA extended primer from 18 to 81 bases long, a total of 64 fragments, was observed. A second synthetic template was designed to yield alternating stretches of dA and dT extensions of the primer. Thirdly, the sequence reaction products from a synthetic oligonucleotide template containing all four bases was analyzed in four independent runs, one for each of the four base-specific reactions. In all cases, the expected number and patterns of peaks were observed by capillary gel electrophoretic analysis. Finally, separation of sequence reaction products generated with single-strand M 13mp 18 phage DNA as template exhibited baseline resolution of fragments differing in length by a single nucleotide and from 18 to greater than 330 bases total length. © 1990.
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收藏
页码:49 / 60
页数:12
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