Colonies not Growing on Plates

[dnamolecule]

Hello,
i am confused.

I have a bunch of bacterial clones growing fine in culture, however when i transfer them to the plates with selection media (penicillin) they dont grow....

any ideas what could be going on?

[aftabac]

well
there are mnay possibilities in this regards
there may be curing of plasmid
your media for plate may not be rich enough to sustain growth of your clone.
when you prepare media then that flask may not be properly cleaned and detergent may be there that stop the growth.
sometime it happens that when you transfer from the growth from broth to plate then it dont grow on it, for this you may have to give more time as access to nutrient is not as much on plate as in broth.
there may be some other reason so you should check carefully one by one that where is problem.
best of luck for you

regards
aftab

[jtendres]

Hello. What aftabac said is true and also, if you're bacterial clones are from a ligation transformation you made, I would check to make sure the vector has the correct insert. What I do to check this is, I do individual plasma preps from ten of my currently growing colonies. Then I do a restriction enzyme digest around the area of my insert, which includes the selection factor, by looking at a plasmid map of restriction enzyme sites. This allows me to run a gel of each DNA to see if the fragments made match each other and using a control that has no insert will allow me to see if it matches that and if so, then my insert is not being added correctly during the ligation.

Hope that helps! Josie