Re: [DIYbio] Re: Reviving bacteria from 1 year old petri dish

Can you miniprep and check plasmid restriction-fragment size, at least?
Or PCR?

On 28/01/15 14:05, Andreas Stuermer wrote:
> Well, we finally got tons of growth of the liquid medium directly plated
> onto a kanamycin agar plate. Without recovery.We put some colonies of
> this into LB kanamycin liquid medium and it grew cloudy.
>
> Unfortunatly we don't have time till the workshop to re-do this from the
> 1 year old agar plate -> LB for recovery -> LB kan for selection.
>
>
> Still, as it is kanamycin resistant I hope it is the correct bacterium
>
>
> On Wed, Jan 28, 2015 at 3:00 PM, Cathal Garvey
> <cathalgarvey@cathalgarvey.me <mailto:cathalgarvey@cathalgarvey.me>> wrote:
>
> I'd let them recover for 30m/1h in SOC or LB, then plate on
> kanamycin, yes. But, even after a year, if they were stable at 4C
> you should be OK.
>
> I've seen 4C in an agar stab recommended as the best way to store
> plasmid-bearing cells, preferable over -20C and possibly even -80C,
> in at least one reputable source. But that's a stab, whereas in an
> overgrown petri dish things might be less stable.
>
> Try your luck, but if it's kanamycin do try a recovery period I think.
>
> On 28/01/15 13:25, Mega [Andreas Stuermer] wrote:
>
> But then you have to breed them for an hour in LB without
> kanamycin first to give them time to produce the enzyme?
>
>
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