DIYbio:Demo

Trying to put together a DIYbio demo in which people transform plasmid DNA to make cells that turn red, glow, or smell like banana's without lab equipment (protocol still uses lab reagents and consumables right now).

1. Thaw cells on ice.
2. Add DNA to cells.
   • Could use either DNA in liquid or ideally spotted onto filter paper.
3. Incubate on ice for 30 minutes.
4. Heat shock at 42°C for 1 minute
   • You can eliminate this step or just microwave water til it gets to 42°C and use that for heat shock. Transformation efficiency improves with heat shock.
5. Recover on ice for 2 minutes
6. Add 300 μL LB media
   • Use LB instead of SOC so you only need one kind of media around
   • Can also add LB+amp media
7. Incubate at 37°C
   • The temperature near the fan of Apple G4's seems to be right at 37°C. So you can just tape samples to the back of a computer. Might want to have a thermometer around to test this cause you don't want it to be too hot and burn the sample. (Stick thermometer into a tube of water taped to back of the computer and let it equilibrate for a bit.) Back of fridges may also work. Suspect that laptops run too hot but haven't checked.
8. Plate on LB-amp plates and incubate at 37°C
   • Tried replacing this with adding to 5mL liquid LB-amp media with mixed success. The luciferase seemed work after two days at room temperature. But the mRFP1 doesn't seems to be as red (though saw a bit of red in cells that settled to the bottom of the tube). The mixed success could be due to the ampicillin in the media going bad after too long at RT?

Notes

1. The mRFP1 turns more red after storage in a fridge for a few hours.
2. The luciferase only works at RT to ~30°C.
3. Need to add isoamyl alcohol substrate to banana.

To do

1. Make more LB media
2. Make more LB-ampicillin plates
3. Miniprep more DNA of BBa_J04450, pTKX15, and BBa_J45200

People

• KK, MC, JB, and RS