超薄平板凝胶中的高速自动化DNA测序

High speed automated DNA sequencing in ultrathin slab gels.

作者信息

Kostichka A J, Marchbanks M L, Brumley R L, Drossman H, Smith L M

机构信息

Department of Chemistry, University of Wisconsin-Madison 53706.

出版信息

Biotechnology (N Y). 1992 Jan;10(1):78-81. doi: 10.1038/nbt0192-78.

DOI:10.1038/nbt0192-78

PMID:1368192

Abstract

We have developed a high speed instrument for automated DNA sequence analysis. The apparatus employs laser excitation and a cooled CCD detector for the parallel detection of up to 18 sets of four fluorescently labeled DNA sequencing reactions during their electrophoretic separation in ultrathin (50-100 microns) denaturing polyacrylamide gels. Four hundred and fifty bases of sequence information is obtained from 100 ng of M13 template DNA in less than one hour, corresponding to an overall instrument throughput of over 8000 bases/hr.

摘要

我们开发了一种用于自动化DNA序列分析的高速仪器。该仪器采用激光激发和冷却的电荷耦合器件（CCD）探测器，在超薄（50 - 100微米）变性聚丙烯酰胺凝胶中进行电泳分离时，可并行检测多达18组四种荧光标记的DNA测序反应。在不到一小时的时间内，从100纳克的M13模板DNA中可获得450个碱基的序列信息，这相当于仪器的总通量超过8000碱基/小时。