Hello,
I was preparing a e coli calcium competent cell (Rosetta Gami strain) for my lab storage. I started with 5 mL of O/N culture diluted in 150 mL of LB with proper antibiotic, but I miss the time and know my culture has OD = 0.9. I was wondering if is it possible to dilute the culture to OD=0.4 with LB medium to reach the proper OD of my protocol?
Many Thanks
It is definitely possible to dilute the culture with the same broth, but for your case diluting the broth won't serve the purpose, For the preparation of competent cells, you need to grow cultures till log phase. Once the log phase has crossed, the cells are no more suitable for transformation, as they are less likely to survive the freeze thaw process.Diluting the culture will reduce the OD by just reducing the cells, but it won't affect the growth phase of the cells.
Therefore, it is recommended to use a culture of OD 0.3-0.5.
-Bhuwan
Dr. Singh thanks for the answer, but those bacs are intended for lab stock, so they'll be used to plasmid and ligations.
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