I am doing a cloning experiment where my vector has the gateway cassette so I need to use SURE 2 cells as they are ccdB immune. However, the protocol for SURE 2 calls for NZY+ broth but our lab does not have this broth and I would prefer to not have to buy the ingredients and make it if I do not have to. My lab does have TB broth, LB broth, and SOC medium however if any of these can be substituted for NZY+ Broth. Thanks for any help.
Another student in our lab went through a whole fiasco with a destination vector. I will share my thoughts. In my opinion all medias should work, but the one they recommend could produce more colonies after plating. Keep in mind that even though your cells are ccdb resistant they grow very slow and produce tiny colonies because they do not like the ccdb cassette. This means you must grow on amp/chl plates and even still there is a high probability to mutate the ccdb casset through natural selection. Therefore I highly recommend picking several colonies and retransforming into non ccdb resistant cells to make sure your plasmids have a functional ccdb casset.
Some medias have extra nutrients to speed up imitial growth for the 1 hr prior to plating.
Thanks for the advice. I may just try the media that has the most similar composition to NZY+ and see how that works out.
Hey Joe,
I believe I just used SOC medium (with the beta-mercaptoethanol from the kit). My controls with PUC18 on ampicillin plates worked fine. My ligations were pretty pitiful though (got like 2 colonies out of 3 plates), but I blamed that on trying to put a 100 bp insert into a 12kb vector.
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