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 .
