Wiemann S, Stegemann J, Grothues D, Bosch A, Estivill X, Schwager C, Zimmermann J, Voss H, Ansorge W
European Molecular Biology Laboratory, Heidelberg, Germany.
Anal Biochem. 1995 Jan 1;224(1):117-21. doi: 10.1006/abio.1995.1015.
We describe an automated DNA-sequencing technique which allows both the simultaneous sequencing from the two strands of double-stranded templates and the subsequent detection of the sequencing products online and in parallel. The technique is based on hardware technology also used in the ALF DNA sequencer (Pharmacia, Uppsala). A helium-neon laser was mounted into the sequencing device additionally to the standard argon laser. Two different primers, labeled with either fluorescein or Texas red, are used in a single sequencing reaction resulting in an output of two sequences. Both sequencing products are then analyzed on-line in the same lanes of a gel. This technique is especially useful for the complete sequencing of DNA fragments up to 1 kb. High accuracy sequencing of PCR products in double-stranded form can now be accomplished in only one sequencing reaction.
我们描述了一种自动化DNA测序技术，该技术既允许从双链模板的两条链同时进行测序，又能在线并行检测测序产物。该技术基于ALF DNA测序仪（Pharmacia，乌普萨拉）中使用的硬件技术。除了标准氩激光外，还在测序设备中安装了氦氖激光。在单个测序反应中使用两种分别标记有荧光素或德克萨斯红的不同引物，从而产生两个序列的输出。然后在凝胶的同一泳道中对两种测序产物进行在线分析。该技术对于完整测序长达1 kb的DNA片段特别有用。现在，仅通过一次测序反应就可以完成双链形式的PCR产物的高精度测序。
