Hey,
I want to check my phages against the bacterial biofilm. I have read papers in which they have added phages in a specific MOI (0.6, 1 etc) either in pretreatment or post treatment of phage infection.
I have a question if cfu/ml of bacteria is1×10^12, it means 1ml of bacterial culture contains 10^12 viable cells. MOI will be calculated accordingly. I don't know the no. of bacterial cells in 100 or 150ul.
How much volume of bacterial culture do you pipette in 96 well microtiter plate? I'm a bit confused as 200ul is the total volume you can pour in 1 well of 96 microtiter plate. How to maintain a specific MOI in each well of microtiter plate?
So long as. you add the same volume of a similar dilution you will have the same number of cells. So figure out how many cells you actually want per well (it probably is much much less that 1x10^12) and then calculate what dilution you need to make so that 0.1ml gives you that number of cells. I'm guessing you probably want something on the order of 1x10^6 or 1x10^7 but of course it depends upon the experiment you are doing.
Reduction of CFU may or may not have any correlation with biofilm reduction, it all depends upon the protocol you are using as to whether you are really reducing biofilm or merely reducing planktonic cells.
No it is not coincidental, it all depends on your protocol.
I agree with @Michael J. Benedik
To enumerate the viable bacteria in a biofilm you need to
1) discard the supernatent,
2) wash with PBS twice
3) resuspend with PBS via scraping with a sterile tip and then the suspension should be diluted and plated.
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