IGEM:Harvard/2010/General Protocols

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 ]]

Plasmid Minipreps: Qiagen Miniprep Kit

To extract plasmid DNA from bacteria, perform a miniprep using the Qiagen miniprep protocol.