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Team:British Columbia/Protocols/Bacteria
From 2011.igem.org
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| + | '''Bacterial Transformation''' | ||
| + | |||
| + | # Thaw 50uL competent cells on ice for 15min | ||
| + | # Add 1-2uL of your plasmid or ligation mix | ||
| + | # Keep on ice for 45min | ||
| + | # Heat shock at 42 degrees Celsius for 45 seconds | ||
| + | # Keep on ice for 2min | ||
| + | # Add 250uL of LB or SOC media | ||
| + | # Incubate (while rotating/shaking) for 1 hour at 37 degrees Celsius | ||
| + | # Spread 50uL and 200uL on selective media plates | ||
| + | # Incubate plates at 37 degrees Celsius overnight and you should see colonies! | ||
Revision as of 19:18, 13 August 2011
iGEM Tips for Success
Things to think about when designing your project and experiments, as well as general safety rules.
Site Directed Mutagenesis
A molecular biology technique in which a mutation is created at a defined site in a DNA molecule.
Bacterial Standard Operating Protocols
How to prepare competent cells, transform your construct into competent cells, and express your protein from a lac promoter.
Yeast Standard Operating Protocols
How to transform your construct into yeast, obtain crude extract for SDS-PAGE, and perform GFP fixation for microscopy and fluorescence-activated cell sorting.
Gas Chromatography-Mass Spectrometry (GC-MS)
A method that combines the features of gas-liquid chromatography and mass spectrometry to identify different substances within a test sample.
Beetle Transfer Experiments
Preliminary experiments to probe the efficiency of transferring yeast via beetle vector.
Yeast-Fungi Co-culture Experiments
Preliminary experiments to investigate the competitive interactions between yeast and the Bluestain Fungus.
Things to think about when designing your project and experiments, as well as general safety rules.
Site Directed Mutagenesis
A molecular biology technique in which a mutation is created at a defined site in a DNA molecule.
Bacterial Standard Operating Protocols
How to prepare competent cells, transform your construct into competent cells, and express your protein from a lac promoter.
Yeast Standard Operating Protocols
How to transform your construct into yeast, obtain crude extract for SDS-PAGE, and perform GFP fixation for microscopy and fluorescence-activated cell sorting.
Gas Chromatography-Mass Spectrometry (GC-MS)
A method that combines the features of gas-liquid chromatography and mass spectrometry to identify different substances within a test sample.
Beetle Transfer Experiments
Preliminary experiments to probe the efficiency of transferring yeast via beetle vector.
Yeast-Fungi Co-culture Experiments
Preliminary experiments to investigate the competitive interactions between yeast and the Bluestain Fungus.
Bacterial Transformation
- Thaw 50uL competent cells on ice for 15min
- Add 1-2uL of your plasmid or ligation mix
- Keep on ice for 45min
- Heat shock at 42 degrees Celsius for 45 seconds
- Keep on ice for 2min
- Add 250uL of LB or SOC media
- Incubate (while rotating/shaking) for 1 hour at 37 degrees Celsius
- Spread 50uL and 200uL on selective media plates
- Incubate plates at 37 degrees Celsius overnight and you should see colonies!
