An improved method of transformation in Pseudomonads |
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Authors: | Shie-Chau Liu Dale A Webster Benjamin C Stark |
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Institution: | (1) Department of Biological, Chemical and Physical Sciences, Illinois Institute of Technology, IIT Center, 60616 Chicago, IL, USA |
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Abstract: | Summary A protocol for producing competent Pseuclomonas aeruginosa, Pseudomonas putida, and Xanthomonas maltophilia was adapted and modified from existing methods. Cells were incubated on ice for 30 minutes in buffered 100 mM MgCl2 followed by 30 minutes in buffered 100 mM CaCl2 prior to addition of DNA. The MgCl2-CaCl2 incubation increased transformation efficiency two to three times, compared with protocols which use incubation in either Mg2+ or Ca2+, but not both. |
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