Searching the web i found that Zymo Research makes a kit for making competent cells and transforming them in aprox 20 seconds without incubating the mix on ice for 30 minutes, without heat-shock at 42 and no SOC recovery. Just mixing the DNA and the competent cells (made with the reagents provided) and plating after the mixing.
Does anybody know if there is a protocol for making the reagents? or any protocol for making competent cells to transform them without heat-shock (nor electroporation)?
Many thanks.
Martin.
I believe the reagent contents are proprietary still but they do give you the protocol to make a whole bunch of tubes of them, so you gan get a lot of cells from their prep kit. If you DNA is in high concentration and you have an amp plasmid then you can add the DNA, incubate 10m on ice, heat shock 42C for 45s, place on ice for 2m and then plate. This quick protocol does not work for kan plasmids though and even the Zymo cells recommend a longer protocol for kan plasmids.
You may want to consult the corresponding patent (US 4981797 A). The composition of the used buffers for this procedure is written suprisingly unambiguous, compared to Qiagens patents.
To me it seems like the DMSO in the transformation buffer does the trick, carrying the DNA through the cell membranes.
Be sure not to use patented information for commercial application.
- Badges
- Science topic
- Similar topics
- Methodology
- Laboratory
- Cellular Biology Techniques