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

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Relation http://ir.cftri.com/12198/
http://dx.doi.org/10.1016/j.mgene.2014.10.002
 
Title CTAB-mediated, single-step preparation of
competent Escherichia coli, Bifidobacteriumsp. and
Kluyveromyces lactis cells
 
Creator Rajagopal, Kammara
Praveen Kumar, Singh
Rajesh, Kumar
Kaneez Fatima, Siddiqui
 
Subject 03 Biochemistry & Molecular Biology
 
Description 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.
 
Date 2014
 
Type Article
PeerReviewed
 
Format application/pdf
 
Language en
 
Rights —
 
Identifier http://ir.cftri.com/12198/1/Meta%20Gene%2C%20Volume%202%2C%20December%202014%2C%20Pages%20807-818.pdf
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.