I would like to grow Mycobact. Smegmatis in 24well plate in TECAN Spark machine. So far, I was using 100ml erlenmeyer flasks with 20 ml bacterial culture w/ shaking 180 rpm in 37C . Tecan would ease my struggle with long experiments (measuring OD every hour for 12 and more hrs). Can anyone tell me if the culture in 24well plate (meaning max 1ml culture/well) grows in the same manner as if you use erlenmeyer flasks and shaker? Can you give me some tips about the settings (orbital shaking-amplitude?, detergent preferred in your experiments' setting....). Also - until now, we used Tween20 to prevent clumping of Mycobacteria.....but I see tyloxapol as a better option. Do you have any good or bad experience with either? Thanks a lot!
Hi Jarmila,
Previous tests that aimed at optimizing bacterial growth conditions of Staphylococcus aureus in another Tecan plate reader (Infinite 200 Pro) showed that with 24 well plates the optimum set of parameter is:
- Half of the maximum well volume (500 ul for the plate you mentioned)
- Shaking in orbital mode at incubation position with an amplitude of 3 mm
You could start optimizing your growth curves on the Tecan Spark using this parameters, further tuning the conditions for your bacteria and keeping in mind that shaking and filling volume critically influence the data quality.
Attached you can find the application note that describes the above mentioned assay optimization performed on the Infinite 200 Pro.
Nicola Vettore thank you very much, this is very helpful! I was wondering exactly which starting volume and amplitude I should try.
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