For molar concentration and related product information, please refer to:
In vitro Cas9 mediated digestion of DNA
1. Add the following components to a sterile, nuclease-free tube on ice:
Components | Volume | Final Concentration | |
sgRNA (300 nM) | 2 µl | ~30 nM | |
Cas9 Nuclease Protein (1000 nM) | 0.60 µl | ~30 nM | |
10X Cas9 Nuclease Reaction Buffer | 2 μl | 1X | |
Nuclease-free H2O | 12.4 μl | - | |
Pre-Incubate for 15 minutes at room temperature | |||
Substrate DNA (30 nM) | 3 μl | 3 nM | |
Total Volume | 20 μl | - | |
2. Collect all components by a brief centrifugation. Incubate the reaction at 37 °C for 1 hour.
3. Analyze fragments by running an agarose gel electrophoresis.
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