Difference between revisions of "Team:Calgary/Experiments"
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<li> Incubate plate at 37°C overnight or until growth is observed </li> | <li> Incubate plate at 37°C overnight or until growth is observed </li> | ||
</ol> | </ol> | ||
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<p> *If transformation fails: </p> | <p> *If transformation fails: </p> | ||
Revision as of 07:29, 3 October 2020
General Wet Lab: E. coli Protocols
TEF1 Promoter Testing
Cellulase Secretion
Omega-3 Production
Biosafety
General Wet Lab: E. coli Protocols
- Mix 100ml of 1X TAE buffer with 1g of regular or LMP agarose (for a 1% gel). For a small gel, mix 30ml of 1X TAE with 0.3g agarose.
- Microwave covered for about 1.5 minutes or until agarose is dissolved and pour into a balanced casting tray. LMP gels should be cast in the fridge.
- Add a volume of Safe-Red dye to be 1/6th the total volume of the sample.
- Run samples at 100V (regular gel) or 80V (LMP gel) until sample is ¾ of the way down the gel.
- Add ≤ 5μl (≤ 1/10 of the cell aliquot amount) of the DNA sample to a chemically competent cell aliquot. Mix by pipetting gently, then incubate on ice for 30 to 45 minutes
- Heat shock at 42°C for 1 minute
- Incubate on ice for 5 minutes
- Add 250μl of plain LB or SOC media aseptically, then incubate for 30 to 90 minutes at 37°C, shaking. *If resistance is Kan, must incubate for at least 1 hour
- Plate 100μl (if big plates) or 50μl (if small or half plates) aseptically
- Incubate plate at 37°C overnight or until growth is observed
*If transformation fails:
- Spin down transformed cells for 5 minutes to pellet
- Resuspend pellet in 100μl of media
- Plate and incubate
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TEF1 Promoter Testing
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Cellulase Secretion
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Omega-3 Production
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Biocontainment
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