How much tme it takes to grow ecoli from glycerol stock?
Answers
Answer:
It depends on both your inoculum size and the final culture volume.
A fairly reproducible procedure is this:
(1) streak plate from -80C glycerol stock.
(2) pick single colonies and inoculate liquid LB (2-3 mL), overnight (10-12 h).
(3) Take a small inoculum (say, 20 uL), and inoculate larger volume of LB (say, 2 mL), in multiple tubes (for a prelim kinetics study)
(4) in about 2.5 to 3 h's time, you should get cells progressing through the log phase (until late log and stationary phases)
(5) introduce a theoretical lag phase here (say 30 min), before first sampling. For this point, cells are presumed to experience little growth.
(6) Monitor the OD by taking a sample in 30 min (or less) interval.
(7) Obtain a growth curve for OD = 0.1, 0.2, 0.4, 0.8, 1.2, 1.5, until saturation point (remember to resuspend well, because cells settle and this distorts OD)
(8) Next time, repeat the same inoculation from step (3) (either maintaining the same culture scale, or upscale it to get approximate results).
Doing a kinetics study would be quite easy (and worthwhile) if you have a Nanodrop for sampling.