SIMULTANEOUS ONLINE DNA-SEQUENCING ON BOTH STRANDS WITH 2 FLUORESCENT DYES

被引:30
作者
WIEMANN, S
STEGEMANN, J
GROTHUES, D
BOSCH, A
ESTIVILL, X
SCHWAGER, C
ZIMMERMANN, J
VOSS, H
ANSORGE, W
机构
[1] EUROPEAN MOLEC BIOL LAB,D-69117 HEIDELBERG,GERMANY
[2] HOSP DURAN & REYNALS,INST REC ONCOL,DEPT MOLEC GENET,BARCELONA,SPAIN
关键词
D O I
10.1006/abio.1995.1015
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We describe an automated DNA-sequencing technique which allows both the simultaneous sequencing from the two strands of double-stranded templates and the subsequent detection of the sequencing products online and in parallel. The technique is based on hardware technology also used in the ALF DNA sequencer (Pharmacia, Uppsala). A helium-neon laser was mounted into the sequencing device additionally to the standard argon laser. Two different primers, labeled with either fluorescein or Texas red, are used in a single sequencing reaction resulting in an output of two sequences. Both sequencing products are then analyzed on-line in the same lanes of a gel. This technique is especially useful for the complete sequencing of DNA fragments up to 1 kb. High accuracy sequencing of PCR products in double-stranded form can now be accomplished in only one sequencing reaction. (C) 1995 Academic Press, Inc.
引用
收藏
页码:117 / 121
页数:5
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