In vitro digestion of DNA with Cas9 Nuclease, S. pyogenes (M0386)
Prepare 300nM sgRNA by diluting the stock with nuclease-free water on ice.
Prepare 30nM substrate DNA with a single target sequence by diluting the stock with nuclease-free water on ice.
Assemble the reaction at room temperature in the following order (total volume 30 µl). 
Pre-incubate for 10 minutes at 37⁰C. 
Add 30nM substrate DNA Mix thoroughly.
Incubate at 37°C for 1 hour.
Proceed with fragment analysis.
