I am currently doing my PhD project which consists of a lot of cloning of new plasmids I am assembling. Our laboratory generally maintains the collection on JM109 strain. But since I am doing a lot of Gibson Assemblies, I have been using electrocompetent DH10B cells for higher efficiency. My question is, can I use standard protocol of preparation of electrocompetent E. coli on JM109 instead of DH10B?
Yes that should work fine. There is nothing special about JM109 that makes it incompatible with the standard protocols used to prepare electrocompetent cells.
Yes, you can adapt the protocol for preparing electrocompetent E. coli cells from DH10B to JM109. However, it's important to note that different strains of E. coli may have slightly different requirements for optimal transformation efficiency, so you may need to optimize the protocol for JM109 cells.
Here's a general outline of how you can adapt the protocol for preparing electrocompetent JM109 cells:
By following this adapted protocol, you should be able to prepare electrocompetent JM109 cells for your Gibson Assembly experiments. It's always a good idea to perform optimization experiments to determine the optimal conditions for your specific application.
As for chemically-competent cells the best will be typical rubidium chloride protocol (many of them in the internet) -it will give you very good results in almost every E. coli (including JM109) if properly done.
Other chemical methods may work with less efficiency.
Alternatively, electroporation protocols will also do nice job.
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