From 2007.igem.org

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• Applications:
  1. Amplification of Biobrick DNA for storage and use.
  2. Transformation of purified plasmids into bacteria

• Time to complete protocol:
  • Lab time: 5min
  • Waiting time: overnight
• Approximate cost of materials: $

Method from primary and secondary reagents

Primary & secondary Reagents Required including controls

• Competent cells (from -70degree freezer)
• DNA for transformation
• LB (cupboard)
• LB-agar plates with selective antibiotic (cool room)

Method including controls

1. Thaw out competent cells by warming in hand.
2. Add 0.5uL resuspended plasmid DNA to 20uL competant cells.
3. Heat shock in water bath at 42 degrees for 30 seconds.
4. Add 200ul LB (flame tip before use).
5. Resuspend cells in LB.
6. Under a bunsen spread resuspended bacteria on Agar Plates with selective antibiotic.
7. Incubate plate overnight at 37 degrees.
8. Place in cold room until needed.

Equipement Required

• 1.7ml microcentrifuge
• Pipettes
• 42 degree water bath (balance room)
• Bunsen burner
• Spreader

References

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