IGEM:Harvard/2010/General Protocols
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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. | 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 | **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 | **Label lid of plate with what you're streaking on it, and your name, and the date | ||
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**Incubate plate overnite at 37˚C | **Incubate plate overnite at 37˚C | ||
| - | + | *Day2: Growing liquid cultures | |
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=Plasmid Minipreps: Qiagen Miniprep Kit= | =Plasmid Minipreps: Qiagen Miniprep Kit= | ||
Revision as of 13:42, 17 May 2010
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.
