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

Heng Zhu | Zendy; Ralph A. Dean | Zendy

Open Access

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

Author(s) -

Heng Zhu,

Ralph A. Dean

Publication year - 1999

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/27.3.910

Subject(s) - biology , bacterial artificial chromosome , escherichia coli , electroporation , plasmid , transformation (genetics) , dna , transformation efficiency , genomic dna , chromosome , genetics , computational biology , genomic library , microbiology and biotechnology , genome , gene , base sequence , agrobacterium

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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