How long do I incubate after transformation? ? - (Sep/02/2008 )

Uhm the TA in the lab told me to incubate for 1h or that even 1.5 hours would be ok. A postdoc just told me that no more than 20min is needed?

Whos right? I thought the bacteria had to recuperate and express resistance genes. Then again since im not selecting Ill increase the amount of bacteria that doesnt contain the plasmid.

To incubate for 60 min or not to incubate? Now THAT is the question

Do you mean the "recovery" step after you have done the heat shock?

For me depends on how picky I want to be. Can put straight onto the plate and incubate overnight if you are in a hurry.
Otherwise I incubate for 30mins-1hour.

The postdoc in my old lab used to say "Its not dodgy if it works!"

I incubate for 1.5 hrs, but I am transforming by electroporation and my bacteria is gram positive, so I am not really sure if that can be different from other kind of bacteria and transformation method.

It depends on the antibiotic. Amp only resistant plasmids can be plated immediately, although I tend to let them recover for an hour. For kanamycin, 1 hour seems to be sufficient. For chloramphenicol and tetracycline, we have found that growth for 1.5 - 2 hours markedly increases the transformation results.

I agree with phage434 - Amp-only I usually either plate immediately or let incubate for 30 minutes. For other antibiotics, I usually go for about an hour.

we usually incubate for 45mins. But I really agree with the 2nd post. Do not be doggy if it works!