title: CTAB-mediated, single-step preparation of competent Escherichia coli, Bifidobacterium sp. and Kluyveromyces lactis cells.
abstract: An efficient and reproducible method for transformation depends on the competency of the organism. We have developed a simple method for the preparation of competent Escherichia coli, Kluyveromyces lactis, and Bifidobacterium sp. by using a buffer containing cetyl trimethyl ammonium bromide (CTAB) and permits efficient uptake of plasmid DNA and ligation-reaction products. Cells are harvested, washed, mixed with 1-10 μg/ml CTAB, incubated, and followed by a buffer wash. For long-term storage of competent cells, bacteria may be frozen in 10% glycerol without the addition of other components. The transformation process is very simple; plasmid DNA and the cells are mixed and incubated for 5-60 min at 4 °C; no heat pulse is required, and the duration of incubation at 4 °C is not crucial.
authors.affiliation: Department of Protein Chemistry and Technology, CSIR-Central Food Technological Research Institute (CSIR-CFTRI), Mysore, India.
authors.forename: Kammara
authors.initials: K
authors.lastname: Rajagopal
authors.affiliation: Department of Protein Chemistry and Technology, CSIR-Central Food Technological Research Institute (CSIR-CFTRI), Mysore, India ; (CSIR-IMTECH) Institute of Microbial Technology, Sector-39a, Chandigarh, India.
authors.forename: Praveen Kumar
authors.initials: PK
authors.lastname: Singh
authors.affiliation: Department of Protein Chemistry and Technology, CSIR-Central Food Technological Research Institute (CSIR-CFTRI), Mysore, India ; (CSIR-IMTECH) Institute of Microbial Technology, Sector-39a, Chandigarh, India.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Kumar
authors.affiliation: Department of Protein Chemistry and Technology, CSIR-Central Food Technological Research Institute (CSIR-CFTRI), Mysore, India ; (CSIR-IMTECH) Institute of Microbial Technology, Sector-39a, Chandigarh, India.
authors.forename: Kaneez Fatima
authors.initials: KF
authors.lastname: Siddiqui
keywords: CTAB; CaCl2 method; Competency; FACS; TSS method of transformation; Transformation