H.influenzae and S.pneumoniae cell culture - H.influenzae and S.pneumoniae grow on plates but not in broth (Oct/03/2008 )
I'm setting up H.influenzae and S.pneumoniae cultures for my downstream experiments. Both strains are "wild type" and have no plasmids. I grow H.influenzae on chocolate agar plates and Mueller-Hinton broth with 15ug/ml hematin, 5mg/ml yeast extract, 15ug/ml beta-NAD. My S.pneumoniae grows on 5% blood agar plates and LB medium. I have no problems growing these bacteria on agar plates (@37C, 5% CO2). However, I cannot establish reproducible growth in broth (sometimes broth inoculation will work and sometimes it won't, however it does happen in cycles). Interestingly, when broth cultures don't grow, both strains are affected.
To the best of my knowledge, I grow these cultures under the "same conditions". Does anybody has an idea of what is happening?
To the best of my knowledge, I grow these cultures under the "same conditions". Does anybody has an idea of what is happening?
"Interestingly, when broth cultures don't grow, both strains are affected."
that really highly implies something is wrong with ur broth prep.
Could u try growing them in just LB or MH without all the fancy stuff?