Literature DB >> 9889291

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

H Zhu1, R A Dean.   

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.

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Year:  1999        PMID： 9889291      PMCID： PMC148265          DOI： 10.1093/nar/27.3.910

Source DB:  PubMed          Journal:  Nucleic Acids Res        ISSN： 0305-1048            Impact factor:   16.971


  6 in total

1.  Role of Escherichia coli O157:H7 virulence factors in colonization at the bovine terminal rectal mucosa.

Authors:  Haiqing Sheng; Ji Youn Lim; Hannah J Knecht; Jie Li; Carolyn J Hovde
Journal:  Infect Immun       Date:  2006-08       Impact factor: 3.441

2.  Construction and characterization of large-insert genomic libraries (BAC and fosmid) from the Ascidian Botryllus schlosseri and initial physical mapping of a histocompatibility locus.

Authors:  Anthony W de Tomaso; Irving L Weissman
Journal:  Mar Biotechnol (NY)       Date:  2003 Mar-Apr       Impact factor: 3.619

3.  Cloning and characterizations of the Serratia marcescens metK and pfs genes involved in AI-2-dependent quorum-sensing system.

Authors:  Hu Zhu; Ya-Ling Shen; Dong-Zhi Wei; Jia-Wen Zhu
Journal:  Mol Cell Biochem       Date:  2008-05-16       Impact factor: 3.396

4.  Direct expression and validation of phage-selected peptide variants in mammalian cells.

Authors:  Brian D Quinlan; Matthew R Gardner; Vinita R Joshi; Jessica J Chiang; Michael Farzan
Journal:  J Biol Chem       Date:  2013-05-10       Impact factor: 5.157

5.  Improved method for high-efficiency electrotransformation of Escherichia coli with the large BAC plasmids.

Authors:  Jana Nováková; Anita Izsáková; Tomáš Grivalský; Christian Ottmann; Marian Farkašovský
Journal:  Folia Microbiol (Praha)       Date:  2013-07-12       Impact factor: 2.099

6.  Genome-scale identification method applied to find cryptic aminoglycoside resistance genes in Pseudomonas aeruginosa.

Authors:  Julie M Struble; Ryan T Gill
Journal:  PLoS One       Date:  2009-11-11       Impact factor: 3.240
  6 in total

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