| Component | Ligation | Control |
| 3ZF GNN vector | 600 ng | 200 ng |
| 3ZF GNN insert | 900 ng | ------- |
| 5× T4 DNA Ligase Buffer | 48 μL | 16 μL |
| T4 DNA Ligase | 12 μL | 4 μL |
| Nuclease-free water | to 240 μL | to 80 μL |
| Total | 240 μL | 80 μL |
| Component | Ligation | Control |
| 3ZF GNN vector | 600 ng | 200 ng |
| 3ZF GNN insert | 900 ng | ------- |
| 5× T4 DNA Ligase Buffer | 48 μL | 16 μL |
| T4 DNA Ligase | 12 μL | 4 μL |
| Nuclease-free water | to 240 μL | to 80 μL |
| Total | 240 μL | 80 μL |