Team:BNDS China/Notebook
Notebook
Q5 PCR Reaction system (50 μL) for the vector RPG
- Q5 Reaction Buffer (5x): 10 μL
- dNTP: 3 μL
- Forward Primer/ Reverse Primer :2.5 μL/2.5 μL
- Template (RPG): 45ng
- Q5 DNA Polymerase: 0.5 μL
- ddH2O：31 μL
Transformation system:
Positive control: 25 μL competent cell + 1 μL RPG
Negative control: 25 μL competent cell
Experiment group (with Gibson products): 50 μL competent cells + 2 μL Gibson products
- Pick 6-10 colonies.
- Add 20 μL Amp+ liquid LB broth into 0.2 mL microfuge tube and add colonies, respectively. Then use 37°C shaker (with rotate speed 270r/min) to revive the bacteria for 1 hour.
Colony PCR Reaction System:
- Taq Mix: 12.5 μL
- Forward Primer/ Reverse Primer: 1 μL/1 μL
- Bacteria LB solution: 2 μL
- ddH2O: 10.5 μL
