For inoculum prep, do we
(a) measure growth at OD600nm and stop when said OD value reaches 0.5. Inoculation volume as set beforehand.
or
(b) do we take a certain amount of culture, and dilute/concentrate if necessary, so that the new inoculated medium's OD is 0.5?
This may seem trivial to some but I'd appreciate any inputs nevertheless. Thank you :)
Hi Mohamad!
The first answer is the right one! You have to measure cell growth at OD600 and stop when OD value reaches 0.5. Initially cells will grow very slowly, but pay attention to measure OD each 10 minutes because when cells will reach the exponential phase they will grow faster.
I hope I have been useful for you!
Silvia
Hi Mohammed
But the second option can also be used if you do a overnight culture before.
Hi Mohammad,
I think the first option is better to go with..! Once you inoculate new media with bacterial inoculum, it takes time to achieve exponential phase/log phase, as the OD is indicator of bacterial growth based on the growth cycle, check OD at set time interval (Varies from 10 to 30 min), and wait until it reaches to 0.5 at 600 nm wavelength.
Second option is good with dilution only if the range of OD is not so wide, I would rather say to stick with the first and more traditional/proven option which is equal to concentrating the growth media upon time.
Vaibhav
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