grow in broth but not on agar... - after transformation (Sep/06/2010 )
Hi, I'd like to ask a question
after i did my transformation, i spread 100ul of the culture on LB agar with appropriate antibiotic, and the remaining i put it in LB broth with appropriate antibiotic
after 16 hours if incubation, i found out there are no colony on the agar plate but there is growth in the broth.
should i spread plate another time on the other plate from the broth?
p/s:sorry for my bad english
-badguy-
you are not suppose to put that 100ul into the LB broth. Even you can see the growth, u cant do anything with that.
When u see the growth on the agar plate, pick some colonies and put those in the LB broth.
-stagius24-
badguy on Tue Sep 7 01:49:20 2010 said:
Hi, I'd like to ask a question
after i did my transformation, i spread 100ul of the culture on LB agar with appropriate antibiotic, and the remaining i put it in LB broth with appropriate antibiotic
after 16 hours if incubation, i found out there are no colony on the agar plate but there is growth in the broth.
should i spread plate another time on the other plate from the broth?
p/s:sorry for my bad english
Instead of spreading 100ul, spin down your LB with bacteria, remove all sup except for ~50ul, resuspend by vortexing 2x4"@3000rpm (with 5" wait in between) and plate everything.
-hematopoietry-