三种使用毛细管凝胶电泳和激光诱导荧光的DNA测序方法。

Three DNA sequencing methods using capillary gel electrophoresis and laser-induced fluorescence.

作者信息

Swerdlow H, Zhang J Z, Chen D Y, Harke H R, Grey R, Wu S L, Dovichi N J, Fuller C

机构信息

Department of Chemistry, University of Alberta, Edmonton, Canada.

出版信息

Anal Chem. 1991 Dec 15;63(24):2835-41. doi: 10.1021/ac00024a006.

DOI:10.1021/ac00024a006

PMID:1789449

Abstract

Capillary gel electrophoresis is demonstrated for the four-spectral-channel sequencing technique of Smith, the two-spectral-channel sequencing technique of Prober, and the one-spectral-channel sequencing technique of Richardson and Tabor. Sequencing rates up to 1000 bases/h are obtained at electric field strengths of 465 V/cm. At lower electric field strengths, capillary electrophoresis produces useful data for fragments greater than 550 nucleotides in length with 2 times better resolution than slab gel electrophoresis. An on-column detector produces detection limits of 200 zmol (1 zmol = 10(-21) mol = 600 molecules) for the four-spectral-channel technique. A postcolumn detector, based on the sheath flow cuvette, produces detection limits of 20 and 2 zmol for the two- and one-spectral-channel techniques, respectively.

摘要

毛细管凝胶电泳已被用于史密斯的四光谱通道测序技术、普罗伯的双光谱通道测序技术以及理查森和泰伯的单光谱通道测序技术。在465 V/cm的电场强度下，测序速率可达1000个碱基/小时。在较低的电场强度下，毛细管电泳对于长度大于550个核苷酸的片段能产生有用的数据，其分辨率比平板凝胶电泳高2倍。对于四光谱通道技术，柱上检测器的检测限为200 zmol（1 zmol = 10^(-21) mol = 600个分子）。基于鞘流比色皿的柱后检测器，对于双光谱通道技术和单光谱通道技术的检测限分别为20 zmol和2 zmol。