Mike Barnkob:Protocols/Bacterial/ON liquid culture
From OpenWetWareJump to navigationJump to search
Inoculating overnight liquid culture
Pick colonies and inoculate a liquid culture.
Prepare liquid LB media:
- Following into a 500 ml flask:
8.2 g LB Broth Ezmix (which includes Tryptone, 5 g/L Yeast Extract, 5 g/L NaCl)
dH2O up to 400mL
- Cover flask with aluminum foil, label with autoclave tape and autoclave.
- Add ampicillin to a concentration of 100 µg/mL (0.4 ml of 100mg/mL)
For a single colony:
- Add 5 ml of LB + antibiotic to two 50ml Falcon tubes.
- Using a sterile loop, select a single colony from LB plate and dip into Falcon Tube.
- Loosely cover the culture with sterile aluminum foil or a cap that is not air tight.
- Incubate overnight (12-18 hours) in shaking incubator at 37°C.
Note: Also incubate a tube that only has LB + antibiotic, but no bacteria as a control. No bacteria should be visible in this after incubation.
- Check for growth
- Proceed to create a glycerol stock and isolate plasmid DNA.