| Method table. Cleavage reaction conditionsa | |||||||
|---|---|---|---|---|---|---|---|
| Figure | Replicates | Cas (nM) | crRNA (nM) | Target | Trans-substrate (nM) | Time (hr) | Other (when indicated) |
| 2A | 3 | 5.0 | 2.5 | 1.0 nM | None | 0.5 | 50 mM EDTA |
| 2B | 3 | 2.0 | 1.0 | 10 pM | 100 | 1.0 | 25 mM EDTA |
| 2C and 2D | 2 | Vary | Vary | 1.0 nM | None | Vary | |
| 2E | 2 | 10 | 5.0 | Vary | None | Vary | |
| 2G and S3C | 1 | 10 | 5.0 | 1.0 nM | Vary | Vary | |
| 3A | 3 | 2.0 | 1.0 | Vary | 10, 100 | 2.0 | |
| 3B and 3C | 3 | 2.0 | 1.0 | Vary | 100 | 1.0 | |
| 3D–3F, S3D, S3E, S3G, and S3H | 1 | 2.0 | 1.0 | 10 pM | Vary | Vary | |
| 3H, 3I, S3F, and S3I | 3 | 2.0 | 1.0 | 10 pM | Vary | Vary | |
| 4A | 3 | 5.0 | 2.5 | 1.0 nM | None | 0.5 | 50 mM EDTA |
| 4B and 6D | 3 | 1.0 | 1.0 | 10 pM | 100 | 1.0 | 50 mM EDTA |
| 4C | 2 | Vary | Vary | 1.0 nM | None | Vary | |
| 4D | 2 | 10 | 5.0 | Vary | None | Vary | |
| 4E | 2 | 5.0 | 5.0 | 1.0 nM | Vary | Vary | |
| 4F | 2 | 5.0 | 5.0 | 1.0 nM | Vary | Vary | |
| 5A, 5B, and 6C | 3 | 1.0 | 1.0 | Vary | 100 | 2.0 | |
| 5C | 3 | 1.0 | 1.0 | Vary | 10, 100 | 2.0 | |
| 5D | 1 | 1.0 | 1.0 | 10 pM | Vary | Vary | |
| 5E | 3 | 1.0 | 1.0 | 10 pM | Vary | Vary | |
| 5F | 1 | 2.0 | 1.0 | 1.0 nM | Vary | Vary | |
| 5G | 1 | 2.0 | 2.0 | 1.0 nM | Vary | Vary | |
| 6A and 6B | 3 | 2.0 | 1.0 | Vary | 100 | 1.0 | |
| 6E and S6A | 4–13 | 1.0 | 1.0 | 10 pM | 100 | 1.0 | |
| 6F | 3 | 1.0 | 1.0 | 10 pM | 10 | 2.0 | |
| 6G | 3 | 1.0 | 1.0 | Vary | 10 | 2.0 | |
| 6H | 3 | 1.0 | 1.0 | Vary | 100 | 2.0 | 0.5 mg/mL BSA |
| S2A | 3 | 2.0 | 1.0 | 100 pM | 83 | 0.5 | |
| S2B | 3 | 1.0 | 0.5 | 500 pM | 100 | 1.0 | |
| S2C | 3 | 2.0 | 1.0 | 10, 100 pM | 100 | 2.0 | |
| S2D–S2H | 3 | 2.0 | 1.0 | Vary | 100 | 1.0 | |
| S3A | 3 | 5.0 | 2.5 | 1.0 nM | None | Vary | |
| S3B | 3 | Vary | Vary | 1.0 nM | None | Vary | |
| S3J | 1 | 8.0 | 4.0 | 10 pM | Vary | Vary | |
| S3K | 1 | 2.0 | 1.0 | 100 pM | Vary | Vary | 100 μg/mL yeast tRNA |
| S4A | 3 | 5.0 | 5.0 | 1.0 nM | None | Vary | 50 mM EDTA |
| S4B | 3 | 10 | 5.0 | 1.0 nM | None | Vary | |
| S4C | 1 | 1.0 | 1.0 | 10 pM | Vary | Vary | |
| S4D | 1 | 4.0 | 4.0 | 10 pM | Vary | Vary | |
| S4E | 1 | 1.0 | 1.0 | 100 pM | Vary | Vary | |
| S4F | 1 | 5.0 | 6.26 | 100 pM | Vary | Vary | 50 μg/mL heparin |
| S4G | 3 | 5.0 | 6.25 | 100 pM | Vary | Vary | 100 μg/mL heparin |
| S5A | 3 | 1.0 | 1.0 | 2.5, 5.0, 10 pM | 100 | 2.0 | |
| S5B | 3 | 1.0 | 1.0 | Vary | 100 | 0.5 | |
| S5C | 3 | 2.0 | 2.0 | 1.0 nM | 83 | 0.25 | |
| S5D | 3 | 1.0 | 1.0 | 1.0 nM | 100 | 1.0 | 50 mM EDTA |
| S5E | 3 | 2.0 | 2.0 | 1.0 nM | 100 | 1.0 | 50 mM EDTA |
| S6B and S6D | 3 | 1.0 | 1.0 | 10 pM | 100 | 1.0 | |
a Cleavage reactions performed in the number of wells for each point in the indicated figures. Concentrations are given for the components during the cleavage phase of the reactions for the time indicated.