Table 1.
The mixture for phosphorylation and annealing of sgRNA oligos
| Top strand for the sgRNA primer | 100 pmol |
| Bottom strand for the sgRNA primer | 100 pmol |
| 10× T4 DNA ligase buffer | 1 μL |
| T4 polynucleotide kinase (PNK) (10 U/ μL) | 0.5 μL |
| ddH2O | Add ddH2O to a total volume of 10 μL |