克隆(编程)
吞吐量
质粒
计算生物学
自动化
协议(科学)
生物
突变
蛋白质表达
功能基因组学
基因
计算机科学
基因组学
基因组
遗传学
突变体
工程类
病理
无线
替代医学
程序设计语言
医学
机械工程
电信
作者
Lynda Dieckman,M. Gu,Lucy Stols,Mark I. Donnelly,F. Collart
标识
DOI:10.1006/prep.2001.1602
摘要
We outline a high throughput process for the production of bacterial expression clones using automated liquid handlers. The protocol consists of a series of interlinked methods representing liquid manipulations or incubations on various stations of the automation system. The methods employ the ligation-independent cloning approach that enables the simultaneous production of plasmids for different expression systems. The current cloning protocol spans 3 days with a linear throughput of 400 targets per production run. This automated approach enables the production of large numbers of bacterial expression clones and ultimately purified proteins. Although they were developed for structural genomics, these molecular protocols can also be applied in high throughput strategies such as those used for site-specific mutagenesis or protein interaction studies.
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