Team:Alberta/Notebook/protocols/overnight
5mL Overnights
Reagents:
- 10mL culture tube Use 16mm x 160mm or 16mm x 125 mm
- 5mL LB
- Concentrated antibiotics
- Single colonies on a plate (Best not to start O/N from glycerol stocks)
Procedure:
- Pipet 1.5-20uL of the appropriate antibiotic to the appropriate working concentration into culture tube.
- Add 5mL non-contaminated LB. Then add antibiotic.
- Select single colony using a sterile instrument that has been cooled and inoculate the liquid culture.
- Place culture tube in incubator at 37oC overnight shaking vigorously (250 rpm).
Notes:
For I0500, culture overnight; then add 1mM ITPG in morning and continue culture for another 3 hours. Then miniprep.