Magni:Protocols/Inocula

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Plasmid propagation

To propagate a plasmid in vivo, infect selective liquid medium with a single colony from a transformation plate.

ALWAYS WORK NEAR THE FLAME!

  • Prepare 5 ml of LB + antibiotic in a 15-ml tube.
  • Select a well-isolated colony from a transformation plate.
  • Take a P20 with a sterile tip.
  • Touch this colony with the tip and infect the liquid medium.
  • Close the 15-ml tube and incubate it at 37degC, 220 rpm overnight (14-24 hours).
  • Seal the transformation plate with parafilm and store in the fridge.

NOTES

  • Heat-sensitive plasmids must be incubated at a proper temperature. If an incubator set at a low temperature is not available, incubate the culture at room temperature on the rotating mixer with rolls.
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