Midiprep -> liquid culture overnight duration

[Mega]

Hi everyone...

 

I just innoculated a liquid culture for a midiprep and relaized that if I let them grow overnight, that would be close to 22 hours...

 

Is that too much?

 

I gave them just 35°C so they may not grow as quick.

 

 

Thanks!

[Mega]

Maybe, just maybe even as low as 30°C would be better...

[Josiah Zayner]

Overnight culture generally has a range of 8-24 hours at least. Once the antibiotic has degraded then the issue becomes selecting for the plasmid. However, I have done preps on cultures that were > 24hours old and there was no issue.

[Andreas Sturm]

Ok thanks.

Did a miniprep of pVIB (because I had no plasmid any more, just frozen bacteria). Also just one colony on the agar plate glowed, the others were all kind of contaminants!! (Maybe it came from unsterilized glycerol. 

result: 179 ng/uL -  800 uL

I let another 500 uL run throught the column/filter and still got 30 ng/uL !! 

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