sporulation in Bacillus

[praveen127]

Can anyone tell me about spores germination in B. pumilus? I am trying to count the sporulation efficiency in my B.pumilus mutants by phase contrast microscopy as well with counting on normal LB agar plate ( after heat shock at 80deg for 20min)..but the results in microscopy showing around 40-50% spores while on plates it shwoing less than 2%. Can anyone tell me the problem? is it due to germination of spores?

[cobaltbomb]

you might want to try to lower your temperature, and or time...
or try to inactivate the vegetative cells with ethanol
maybe these spores are not very heat resistant, or they germinate prior, or during your heating, wich would kill them.
waht mutation is it
85°for 15 minutes incubated in wet heat (in the medium) killed clostridium spores when i tried it