USE OF NON-CROSS-LINKED POLYACRYLAMIDE FOR 4-COLOR DNA-SEQUENCING BY CAPILLARY ELECTROPHORESIS SEPARATION OF FRAGMENTS UP TO 640 BASES IN LENGTH IN 2 HOURS

被引：104

作者：

ZHANG, JZ ^{[1
]}

FANG, Y ^{[1
]}

HOU, JY ^{[1
]}

REN, HJ ^{[1
]}

JIANG, R ^{[1
]}

ROOS, P ^{[1
]}

DOVICHI, NJ ^{[1
]}

机构：

[1] UNIV ALBERTA, DEPT CHEM, EDMONTON, AB T6G 2G2, CANADA

来源：

ANALYTICAL CHEMISTRY | 1995年 / 67卷 / 24期

关键词：

D O I：

10.1021/ac00120a026

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Four-color DNA cycle sequencing was performed on an M13mp18 template using dye-labeled primers, Sequencing fragments were separated by capillary electrophoresis at 60 degrees C and at an electric field of 150 V/cm. The sieving medium was 5%T, non-cross-linked polyacrylamide in 7 M urea. The use of high temperature for the separation reduces formation of secondary structures in the sequencing fragments, generating a sequence that is free of compressions without the use of strongly denaturing gels. The use of high temperatures also increases the separation rate compared with room-temperature operation, Fragments up to 640 bases are separated in less than 2 h.

引用

页码：4589 / 4593

页数：5

共 27 条

[1] DNA SEQUENCING - PRESENT LIMITATIONS AND PROSPECTS FOR THE FUTURE [J].