I have bacterial culture of BL21 DE3 transformed with pET28a containing the gene of interest, stored in 4 degree Celsius from 4 months. Do i use culture stored at 4 degree or glycerol stock stored at -80 degree Celsius?
Protein expression efficiency is the main concern?
Hi
You should not use old transformed culture more than one month old because it is possible that the loss of plasmids occurs. You should try with fresh inoculation or revived culture.
The glycerol stocks from -80C work better. I would not use the cultures from 4C if they are stored more than 3 weeks. Good luck
Dear friends, I have a question in this around. To store E.coli at -20 or - 80 C, is it required to flash-freeze the sample in liquid nitrogen before storing? Thanks
it´s not necessary, you can make a transition from -20° to -80° with the culture in the broth + glycerol.
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