一种提高大肠杆菌转化效率的新方法——用于细菌人工染色体文库构建
A novel method for increasing the transformation efficiency of Escherichia coli-application forbacterial artificial chromosome library construction.
作者信息
Zhu H, Dean R A
机构信息
Clemson University Genomics Institute, Clemson, SC 29634, USA.
出版信息
Nucleic Acids Res. 1999 Feb 1;27(3):910-1. doi: 10.1093/nar/27.3.910.
Abstract
Bacterial artificial chromosome (BAC) libraries play a pivotal role in genomics studies. A crucial step in BAC library construction is the transformation of Escherichia coli by electroporation. Absolute efficiency (cfu/microgram DNA) is affected by a number of factors including the topological form and treatment of DNA samples. Here we report a simple new protocol using tRNA assisted precipitation that increased transformation efficiency by 70-fold for BAC ligations and up to 400-fold for plasmid ligations. The mechanism may involve altering or stabilizing the topographical form of the DNA molecules.
摘要
细菌人工染色体（BAC）文库在基因组学研究中发挥着关键作用。BAC文库构建的一个关键步骤是通过电穿孔法转化大肠杆菌。绝对效率（每微克DNA的菌落形成单位）受多种因素影响，包括DNA样品的拓扑形式和处理方式。在此，我们报告一种使用tRNA辅助沉淀的简单新方法，该方法使BAC连接的转化效率提高了70倍，质粒连接的转化效率提高了多达400倍。其机制可能涉及改变或稳定DNA分子的拓扑形式。
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