Pohl T M, Maier E
GATC Gesellschaft für Analyse-Technik und Consulting mbH, Konstanz, Germany.
Biotechniques. 1995 Sep;19(3):482-6.
We have developed efficient protocols for the construction of shotgun libraries and for high-throughput DNA sequencing at low cost using the optimized GATC 1500 Direct Blotting Electrophoresis system with colorimetric detection on nylon membranes. We can now sequence entire cosmids without any primer walking at low coverage (8-fold to 9-fold). We have demonstrated the efficiency of our protocols while sequencing 500 kb of yeast DNA with the GATC 1500 system. The methods described here have allowed us to achieve the highest data rate within the framework of the European Communities yeast sequencing program. In 1993, only two people were involved in generating and submitting 156,274 bp of final sequence data. Thereof, 39,360 bp have been verified independently in other laboratories, and no sequencing error has been found.
我们已经开发出了高效的方案，用于构建鸟枪法文库以及使用优化的GATC 1500直接印迹电泳系统在尼龙膜上进行比色检测，以低成本进行高通量DNA测序。我们现在可以在低覆盖度（8倍至9倍）下对完整的黏粒进行测序，而无需任何引物步移。我们已经在使用GATC 1500系统对500 kb酵母DNA进行测序时证明了我们方案的有效性。这里描述的方法使我们能够在欧洲共同体酵母测序计划的框架内实现最高的数据率。1993年，只有两个人参与生成并提交了156,274 bp的最终序列数据。其中，39,360 bp已在其他实验室中独立验证，未发现测序错误。
