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

Heng Zhu; Ralph A. Dean

doi:10.1093/nar/27.3.910

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

Heng Zhu, Ralph A. Dean

Research output: Contribution to journal › Article › peer-review

18 Scopus citations

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/μg 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.

Original language	English (US)
Pages (from-to)	910-911
Number of pages	2
Journal	Nucleic acids research
Volume	27
Issue number	3
DOIs	https://doi.org/10.1093/nar/27.3.910
State	Published - Feb 1 1999
Externally published	Yes

ASJC Scopus subject areas

Genetics

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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/μg 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.",

author = "Heng Zhu and Dean, {Ralph A.}",

note = "Funding Information: This is technical contribution no. 4457 of the South Carolina Agricultural Experiment Station. This research was supported in part by National Science Foundation Grants nos IBN951308 and DBI9724557, and Clemson University.",

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N2 - 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/μg 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.

AB - 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/μg 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.

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

Abstract

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