Golden Gate
- Prepare the reaction (20 uL volume):
- Add 40 fmol/uL of the following:
- Each insert
- Plasmid
- 2 uL of T4 ligase buffer (10X)
- 2 uL of BSA (10X)
- 0.5 uL of 400,000 U/mL T4 ligase
- 0.5 uL of BsaI-HF
- Water to 20 uL
- Run in a thermocycler for 30 cycles between 30°C (2 min) and 16°C (2min), with 2 holds at 50°C (5 min) and 80°C (5 min) to heat inactivate enzymes
