16 replies to this topic

[Tan Mei Yin]

dear pito and El Crazy Xabi

I've decided to change the MRS broth's pH to 8.36 (the pH level where bile dissolved completely in.). And the control will be 0% bile (which is just only normal MRS broth).
I think the problem with bile being clumped together after autoclave depends on the brand (the ingredients and the overall pH level of the broth too) of MRS that you're using. My supervisor said there's no problem with the one with Oxoid (sadly we couldn't get Oxoid's).

I've done the experiment on it and it seemed okay, the bacteria that was assumed to be lacto did grow in the 0.2% bile concentration. (i did it according to this journal).

Unfortunately there's another problem..we think that the spectro is not working well so we're using microplates instead. To save the broth usage too.

So..could I ask another question? I'm not sure how much microlitre for 1 well in the microplate needs. If i assume that it needs 1microlitres, is it fine if i inoculate 1% of the bacteria into it? (1% of inoculant in 1 microlits. broth = 10 microlits.) or 10%?

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[pito]

Tan Mei Yin, on 26 October 2012 - 10:05 PM, said:

dear pito and El Crazy Xabi

I've decided to change the MRS broth's pH to 8.36 (the pH level where bile dissolved completely in.). And the control will be 0% bile (which is just only normal MRS broth).
I think the problem with bile being clumped together after autoclave depends on the brand (the ingredients and the overall pH level of the broth too) of MRS that you're using. My supervisor said there's no problem with the one with Oxoid (sadly we couldn't get Oxoid's).

I've done the experiment on it and it seemed okay, the bacteria that was assumed to be lacto did grow in the 0.2% bile concentration. (i did it according to this journal).

Unfortunately there's another problem..we think that the spectro is not working well so we're using microplates instead. To save the broth usage too.

So..could I ask another question? I'm not sure how much microlitre for 1 well in the microplate needs. If i assume that it needs 1microlitres, is it fine if i inoculate 1% of the bacteria into it? (1% of inoculant in 1 microlits. broth = 10 microlits.) or 10%?

I am not sure I understand you.
You cant to fill each will of your microplate with 1µl of cellsuspension?
Thats not a lot...

I am not sure what you are trying to do really..

You can use a microplate (the microwells) as wells to grow your bacteria in: you just add a certain volume of broth and inoculate it with a single colony of you bacterium from a plate (for example)