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From 2010.igem.org

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Monday, July 5 2010

Transformation of XL1-B with ligations (failed)

Followed competent cell making protocol (quick CaCl2 method).

Had a small but visible pellet after centrifuging.

Started liquid culture of Gary’s XL1-Blues, added 1 mL of LB and incubated.

• 12 µl pGEM ligation
• 12 µl pNoTat ligation
• 5 µl RFP control DNA

Separate tubes, added our competent cells to each.

   * Incubated for 20-30 minutes on ice (25 min).
   * Heat shock 2 minutes at 42°C (actual: 44-45°C, don’t know if this affects anything.
   * Plated 50 µl out on each amp++ plate

20 µl on each 1/3 of null plate.

Incubated at 37°C overnight; start at 7:15 PM.