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