Is it essential to incubate shigella for 18-24 hrs before starting the procedure of outer membrane proteins separation? What happens if the incubation period increased for more than 24 hours? Also, what happens if the 24-hours bacterial culture was stored in the deep freezer (without subjection to ultrasonication) for the next day and we started the separation procedure then?
It is better to have a culture with minimum of dead cells, so early stationery phase is the best. So incubating longer would have many dead cells will contaminate your OMV preps.
1. When you say incubation, I'm assuming you are incubating with aeration. For Shigella, I would incubate in the range of 16-18 hrs.
2. Are you overexpressing a membrane protein or just growing it up and then doing membrane fractionation?
3. I would use fresh culture to do any membrane preps.
just my 2cents.
EK
Best quality of cultures is obtained after 18 hrs incubation; if you wish to store long term, than -80 Celsius on cryo-beads is best (2-3 yrs). If you wish to store medium term (
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