Team:Brown/Notebook/July5

From 2010.igem.org

Revision as of 22:17, 19 July 2010 by Tgjohnst (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

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.