使用通过六聚体连接组装的引物进行自动化四色DNA测序。

Automated four-color DNA sequencing using primers assembled by hexamer ligation.

作者信息

Kaczorowski T, Szybalski W

机构信息

McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, Madison 53706, USA.

出版信息

Gene. 1996 Nov 7;179(1):195-8. doi: 10.1016/s0378-1119(96)00325-3.

DOI:10.1016/s0378-1119(96)00325-3

PMID:8955647

Abstract

A procedure based on the assembly of sequencing primers by hexamer ligation and then using them in automated DNA sequencing is described. This method is based on a four-color fluorescent terminator chemistry. Sequencing ladders were analyzed using an ABI 373 DNA sequencer (Applied Biosystems, Foster City, CA, USA). The best results were obtained for primers assembled by ligation of four to ten hexamers. The accuracy of the method was estimated to be 99.5% up to 400 nt of the read sequence, and somewhat lower at 400-600 nt.

摘要

描述了一种基于通过六聚体连接组装测序引物，然后将其用于自动DNA测序的方法。该方法基于四色荧光终止剂化学。使用ABI 373 DNA测序仪（美国应用生物系统公司，加利福尼亚州福斯特城）分析测序梯。通过连接四至十个六聚体组装的引物获得了最佳结果。该方法的准确性估计在读取序列的400 nt以内为99.5%，在400 - 600 nt时略低。