Open AccessA 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.