SIMULTANEOUS MONITORING OF DNA FRAGMENTS SEPARATED BY ELECTROPHORESIS IN A MULTIPLEXED ARRAY OF 100 CAPILLARIES

被引:199
作者
UENO, K
YEUNG, ES
机构
[1] US DOE,AMES LAB,AMES,IA 50011
[2] IOWA STATE UNIV SCI & TECHNOL,DEPT CHEM,AMES,IA 50011
关键词
D O I
10.1021/ac00081a010
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Various excitation schemes for distributing a laser beam to a large number of capillaries in an array are evaluated. This led to the construction of a multiplexed system for monitoring the electrophoresis of DNA fragments in 100 capillaries. The laser excited fluorescence signals from each capillary are simultaneously recorded at the rate of 0.6 frame/s by a CCD camera. The reconstructed electropherograms show excellent reproducibility and minimal cross-talk. The system provides for two simultaneous excitation wavelengths so that it can be adapted for two-color, two-intensity DNA sequencing based on the commercial four-dye chemistry. Only 20 mW per laser line was employed. Further development of this system to accommodate up to 4096 independent sequencing channels at a time is discussed.
引用
收藏
页码:1424 / 1431
页数:8
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