Growing a Bacterial E. coli Culture
E. coli is a typical bacteria used for molecular cloning. Long-term storage of BioBrick plasmids is in E. coli suspended in glycerol (500ul cells:500ul glycerol) and kept at -80˚C. Growing E. coli is performed at 37˚C, at which temperature they divide every 20 minutes.
- Day1: Making streaks from glycerol stocks
- Warm agar plate at 37˚C for 10 minutes
- Label lid of plate with what you're streaking on it, and your name, and the date
- Locate desired glycerol stock in -80˚C freezer and, using a sterile toothpick, scrape out a tiny bit and streak on plate
- Incubate plate overnite at 37˚C
- Day2: Growing liquid cultures
- Label 15mL falcon tube and add 5mL of LB media + selective antibiotic
- Using 200uL tip w/o filter, touch streak or single colony and put tip in falcon tube
- Grow culture overnite in a shaking 37˚C incubator
Plasmid Minipreps: Qiagen Miniprep Kit
To extract plasmid DNA from bacteria, perform a miniprep using the Qiagen miniprep protocol.