preparation of LB/ampicillin/IPTG/X-gal plate
Posted 08 April 2011 - 07:49 AM
for preparation of LAIX (LB/Ampicillin/IPTG/X-Gal) plates, i follow this protocol:
Take LB / amp plates into laminar flow, spread 100 ul IPTG onto plates and let adsorb by placing inverted in 37 deg C incubator for 30 min. After this, spread 20 ul of X-Gal onto plates, then let adsorb again at 37 deg C.
Now, the problem is, after taking the LB-ampicillin plate out of the fridge, i did not warm it first
so there was a lot of moisture on the surface of the plate
i proceed directly to spreading IPTG --> incubation --> spreading x-gal --> incubation.
after that, when i looked back at the plate, they were all wet.
and it is only at this time that i realize that i forgot to dry the plates after taking them out of the fridge.
so can my plates be used now?
can anyone help?
Posted 08 April 2011 - 08:47 AM
Posted 08 April 2011 - 11:50 AM
I know that the changes are small with the antibitiocs on the plate but still, cant you get contamination?
I leave my plates to dry in the laminair flow to avoid contamination and yes, they might be not 100% dry.. but it works for me.
Posted 12 April 2011 - 06:54 PM
You could also leave the plate with its over open beside a bunsen burner flame. The flame will keep things from settling on the plate.
People working with laminar flow hoods tend to be a protective when you suggest bring in a plate of bacteria.