Table 3. In vitro transcription reaction for synthesis of sgRNA.
| Reagent | Volume |
|---|---|
| 5× Transcription buffer | 4 μL |
| ATP/GTP/CTP/UTP, 10 mM each | 4 μL |
| DNA template | 400 ng |
| RNase inhibitor | 0.5 μL |
| T7 RNA polymerase | 1 μL |
| DNase/RNase-free H2O | to 20 μL |
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