A novel method for increasing the transformation efficiency of Escherichia coli -: application for bacterial artificial chromosome library construction

被引：20
作者：
Zhu, H [1 ]
Dean, RA [1 ]
机构：
[1] Clemson Univ, Genom Inst, Clemson, SC 29634 USA
来源：
NUCLEIC ACIDS RESEARCH | 1999年 / 27卷 / 03期
基金：
美国国家科学基金会;
关键词：
D O I：
10.1093/nar/27.3.910
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
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/mu g 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.
引用
收藏
页码：910 / 911
页数：2
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