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grow in broth but not on agar...


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#1 badguy

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Posted 06 September 2010 - 05:49 PM

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

#2 stagius24

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Posted 06 September 2010 - 07:55 PM

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.

#3 hematopoietry

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Posted 07 September 2010 - 11:40 PM

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.




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