CTAB-mediated, single-step preparation of competent Escherichia coli, Bifidobacteriumsp. and Kluyveromyces lactis cells

Rajagopal, Kammara and Praveen Kumar, Singh and Rajesh, Kumar and Kaneez Fatima, Siddiqui (2014) CTAB-mediated, single-step preparation of competent Escherichia coli, Bifidobacteriumsp. and Kluyveromyces lactis cells. Meta Gene, 2. pp. 807-818.

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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% glycerolwithout 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.

Item Type:	Article
Uncontrolled Keywords:	Transformation CTAB TSS method of transformation CaCl2 method Competency FACS
Subjects:	500 Natural Sciences and Mathematics > 07 Life Sciences > 03 Biochemistry & Molecular Biology
Divisions:	Protein Chemistry and Technology
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	20 May 2016 10:43
Last Modified:	20 May 2016 10:43
URI:	http://ir.cftri.res.in/id/eprint/12198

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