IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/06/25

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Week 1

Competent Cells

  • Put 50mL LB with 1 mL of dH10B (2 500 ml cylinders) in the shaker at 9:23 am. (Vallari Somayaji)
  • OD of the flasks were checked at 1:00 p.m. The OD was found to be about 1.4. This is concentration is too high to make competent cells. Because of time constraints, a sample was transferred to culture tube containing 5 ml of LB. New flasks will be made on 6/26 to try again.

Transforming cells

  • The following parts were picked from the iGEM registry plates:
Tube Registry Number
1K654058 - TesA
2E0240 - GFP Generator
3J06702 - mCherry
4R0040 - TetR Promoter
5R0011 - LacI Promoter
6B0034 - RBS
7B0032 - RBS
  • Each part was re-hydrated in 10 μl of water.
  • 30 μl of competent cells, donated from the Haynes lab, was added to each DNA sample.
  • The 7 samples were left on ice for 30 minutes.
  • Heat shocked at 42°C for 35 seconds.
  • Placed back on ice for 2 minutes
  • 750 μl of LB media was added to each tube
  • Tubes were placed in shaking incubator for 1 hour at 32°C
  • 350 μl of supernatant was removed from each tube
  • Pellets were re-suspended and placed onto LB agar plates with chloramphenicol.
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