IGEM:Harvard/2010/General Protocols: Difference between revisions
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=Growing a Bacterial E. coli Culture= | =Growing a Bacterial E. coli Culture= | ||
E. coli is a typical bacteria used for molecular cloning. Long-term storage of BioBrick plasmids is | 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= | =Plasmid Minipreps: Qiagen Miniprep Kit= | ||
Revision as of 10:42, 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.
