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Factors which influence CaCl 2 dependent transfection of Lambda DNA in Escherichia coli K12 recipients
Author(s) -
Pfeifer M.,
Pöhlmann Ch.,
Betcke A.,
Kurth M.,
Liebscher D.H.
Publication year - 1980
Publication title -
zeitschrift für allgemeine mikrobiologie
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0044-2208
DOI - 10.1002/jobm.19800200406
Subject(s) - escherichia coli , lambda , transfection , dna , lambda phage , chemistry , genetics , microbiology and biotechnology , biology , physics , gene , bacteriophage , optics
This paper presents further parameters influencing the competence, the process of DNA uptake and the efficiency of plating of CaCl 2 ‐treated E. coli K12 strains. We have found that the process of DNA uptake depends not only on the treatment of bacteria with a certain CaCl 2 ‐concentration but is also influenced considerably by a shift‐down of the CaCl 2 ‐concentration in the reaction mixture. The pH of the growth media and of the reaction mixture plays an important role in maintaining of optimal transfection. The efficiency of plating is influenced by the thickness of the top layer and the concentration of bacteria on the plate. Without genetic variation of the strains, by only varying the mentioned factors we could improve the efficiency of CaCl 2 transfection at about two orders of magnitude to a maximum of 6 × 10 5 pfu/μ DNA.