I have isolated two bacillus bacteria, I want to draw a growth curve for then, please any one can tell me how I can perform this experiment efficiently,
I want two know the following thing
How can I manage the experiment efficiently?
How many replications are required?
Either I need to perform excrement in conical flask or test tubes etc.?
If someone explains the excrement in detail I’ll be very grateful
You have to do according to the following steps:
-Transfer single colony in 250 ml flask contains 100 ml media; grown the culture for 24 hours 130- 150 rpm and temp 37.
-For each isolate you ,made 3 flasks so you will perform experiment in triplicate.
-Next day transfer 1 ml inoculum from 24 hour grown culture to all these 3 flasks.
-Transfer the flasks in the shaker with the conditions mentioned above.
-After transferring it , withdrawn 1-2 ml sample at every hour or 2 hour interval aseptically in LAF.
-OD will be measured at 600 nm.
-Do this kind of withdrawal of sample until you will get constant OD for the culture.
-Plot the graph Hours (X- axis) vs OD (y axis).
-Best of luck.
PS please make sure you know the difference between excrement and experiment
I am very thankful to Traci L Testerman and Parth Thakor for guiding me but I need a little more guideline. if I culture the bacteria in flasks than how can I avoid the contamination while taking the sample at one hour interval?
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