Haynes Lab:Notebook/Synthetic Chromatin for Cancer Research/2015/11/09

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  • Ligation from 11/6 failed. You CANNOT use the classic ligation for beginners. You need to follow the long transformation and heat shock the ligation.
  • Protocol for ligation:
  1. 5ul of ligation & 50ul of cells
  2. Ice for 5 min
  3. Incubate at 42 degrees Celsius for 45 seconds
  4. Ice for 5 min
  5. Add 300ul of SOC
  6. Incubate in the shaking incubator at 37 degrees Celsius for 30-40 min
  7. Spin @ 1000 RCF, decant supernatant
  8. Resuspend cells and plate