Hello,
I need to grow E. coli in 384 microplates directly from colonies. They contain an Amp resistance plasmid. We cannot shake the plate. I tested LB, but they grow poorly. Any suggestion to increase the final OD? Thanks for all thoughts.
Dear Mahmoud
E.coli require oxygenation to growth up to high cell density, therefore i think that you have to find a way for oxygenate you plate and shake it seems the simple way with so small volumes.
If you would like you can replace LB with media for high cell density as Enpresso or YGR (you can find information about it in my blog ProteoCool ( https://proteocool.blogspot.com/ ) but also with this culture media you will need to find a way to mix and oxygenate the culture.
In larger volume a possible alternative is the direct oxygenation by flux the air inside the flask but in a so small scale i think it is very difficult.
A possible alternative is just replicate the colonies in one other solid agar plate, let it to growth a lot and use this for your purpose.
Is plasmid extraction the next step? Colony PCR screening?
Best regards
Manuele
Manuele Martinelli thanks for your advise. We use the plate as a seed culture for protein expression in 96 well format. I will give your suggestion a try. Thanks
Dirk Schmidt Thanks Dirk. I also think testing different media is a good idea.
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