DIYbio:Demo

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Contents

Image:TransformationFoo-v1.png

detailed instructions

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. Punch out DNA sample from part datasheet.
  3. Add punched out paper to cells.
  4. Incubate cells plus DNA on ice for 30 minutes.
  5. Microwave a bowl of water.
  6. Let cool to 42°C
  7. Dip tubes in water for 1 minute
    • You can eliminate this step. Transformation efficiency improves with this step.
  8. Put cells back on ice for 2 minutes
  9. Add 500 μL LB media
    • Use LB instead of SOC so you only need one kind of media around
    • Can also add LB+amp media
  10. Put cells + media in a warm place (ideally 37°C) for 1 hour. 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.
  11. If you are doing red or luciferase, dump onto a plate and spread around by tilting plate.
    • If you are doing banana, add to 5mL LB + antibiotic.
      • The luciferase seemed work after two days at room temperature.
      • But the mRFP1 works far better on a plate than growing in liquid LB-amp.
  12. Put in a warm place (back of a computer etc.)

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
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