I'm trying to transform Bacillus subtilis to fluorescence, and I will use this IGEM protocol with medium MNGE, but there's a part that I can't understand.
It says: "Measure the OD600 of your overnight culture and dilute it in MNGE
Medium to an OD600 of around 0.1-0.2/ml in 10 ml LB"
I don't understand if I should dilute and grow the bacterias in MNGE or dilute and MNGE, after in LB and grow it in LB. If someone helps, I will be thankful.
Hi Clara. I am using that protocol and works fine.
You must dilute your O.N culture in MNGE medium until it reach and O.D of 0.1 OD. In our experiments we use 500 uL of overnight culture in 5 ml of MNGE medium. Once you dilute your O.N culture you let the grow in MNGE medium until OD600 1.0-1.2.
In my experience works very well
Hope that helps.
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