Why does DH5alpha not grow for Inoue competency at 18deg?

05 November 2018 0 10K Report

I am trying to prepare the 200mL batch of Ecoli to make them competent using the Inoue protocol.

The first time I did it, I had no problem to grow a batch of TOP10 cells, just according to the protocol.

However, the DH5alpha strain does not grow at all at 18deg, 200rpm overnight. Last time I had to grow the DH5alpha at 30degreesC in the morning so that I could do the procedure. On the other hand, this batch did result in pretty good competency (about 10to7).

Should I continue doing it in this manner or do you have any suggestions why the DH5 does not grow at 18degrees?

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