Team:Paris/September 9

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Checking our ligases (following)

Ligation (2 hours at 16°C)

  • 4 µL DNA
  • 2 µL 10X buffer 2
  • 1 µL T4 DNA ligase
  • 13 µL water

Transformation

  • Strain used: Chemically competent E. coli DH5 alpha cells
  • 100 µL of cells
  • 5 µL of ligation products
  • 30 min on ice
  • heat shock: 45 s at 37°C
  • 2 min on ice
  • 900 µL of SOC
  • 1 hour at 37°C
  • plating (1 µL or 100 µL) on LB plates containing ampicilline