Adding L-glutamine to DMEM media w/o L-glutamine - (Jul/26/2010 )
I'm trying to do an MTT assay using no phenol-red, no L-glutamine DMEM media and will be transferring mesenchymal stem cells from (+)phenol red (+) L glutamine media into this no L-glutamine media, no phenol-red media .
I was wondering, how much L-glutamine should I add into the no-L glutamine DMEM? There is 500 ml of media.
I think you want a final concentration of 10 mM.
I'm working with mouse bone marrow cells and the basal media that I use (RPMI 1640 - Invitrogen) that already has L-Glutamine in is at a final concentration of 2mM. Just my two cents!
Actually, yes, 2 mM is more what I recall. The bottles of medium with L-glut in them should have the concentration on the label or product sheet.
does adding extra 2mM of glutamine to the media have any adverse effect ( if the protocol requires only 2mM )?
It could do, I'm not sure. I guess what the manufactorer provides in the media is a kind of standard/basal amount of L-Glutamine. The best way to figure this out for your cells would be to try adding extra and not adding extra L-Glutamine to your media and having a go at culturing your cells. It's a small factor but the smallest of things can sometimes have the biggest impacts!