The OD of the bacterial culture for Storing glycerol stocks is 0.8 and after how many hours does the OD reaches 0.8? Kindly help
You should make a growth curve of E.coli through which you get an idea of how much time E.coli OD reaches 0.8. Besides that, it also helps to plan your future experiment related to E.coli culture.
Both answers are good, I would add that in every E.coli culture there's always a lag phase from 0.5 to 1 hour where the O.D doesn't get up this is the latency phase only after this period bacteria are starting to divide. So to answer properly your question all depends of several factors
- how much bacteria you add to the liquid culture
- the volume of the liquid culture
- source of your input bacteria plates or frozen stocks
- the temperature of the culture (works better if you prewarm your medium before inoculation)
The time it takes for E. coli to reach O.D. 0.8 depends on several factors. Inoculate 5-10ml of LB media with 1% culture (or from a single colony on an agar plate). Incubate overnight (12-16hrs) at 37deg C. Add 500 μL of the overnight culture to 500 μL of 50% glycerol (autoclaved).
The duration it takes to reach an OD600 of 0.8 depends on many factors:
- the volume ratio of the inoculated bacteria to the culture medium
- the nature and size of the gene of interest as well as the plasmid carrying it
- the culture temperature
- the number of plasmids co-transformed (in the case of multiple plasmids), hence the number of antibiotics
- the concentration of the antibiotic used
- the strain of the bacteria used, etc.
However, for a typical bacterial strain inoculated into the culture medium at 1:100, and cultured at 37 deg, the OD600 should get to 0.8 within 2.5-3.5 hrs...
But if you want to make a glycerol stock, the ideal thing is to add 500ul of an overnight (12-16 hours) culture to 500ul of 30-50% glycerol.
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