A multiple-capillary electrophoresis system for small-scale DNA sequencing and analysis |
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| Authors: | Zhang J Voss K O Shaw D F Roos K P Lewis D F Yan J Jiang R Ren H Hou J Y Fang Y Puyang X Ahmadzadeh H Dovichi N J |
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| Institution: | Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada. |
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| Abstract: | A five-capillary system has been developed for DNA sequencing and analysis. The post-column fluorescence detector is based on a sheath-flow cuvette. The instrument provides uniform and continuous illumination of the samples. The cuvette virtually eliminates cross-talk in the fluorescence signal between capillaries. Discrete single-photon counting avalanche photodiodes provide high efficiency light detection. The instrument has detection limits (3sigma) of 130 +/- 30 fluorescein molecules injected onto each capillary. Over 650 bases of sequence at 98.8% accuracy were generated in 100 min at 50 degrees C from M13mp18. Separation and detection of short tandem repeats proved efficient and accurate with the use of internal standards for direct comparison of migration times between capillaries. |
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