miniPCR bio
TM
Chopped! Using CRISPR/Cas9 to cut DNA - Student's Guide
Version: 1.0 - Release: May 2022 - © 2022 by miniPCR bio™
Student's Guide
P./22
3. Thoroughly mix the reagents by pipetting up and down 10 times, then cap the tubes
• If you have a vortexer, you can mix the reagents by vortexing for 5 seconds.
4. Make sure all the liquid volume collects at the bottom of the tube
• If necessary, spin the tubes briefly using a microcentrifuge or shake down with a flick of the
wrist.
5. Incubate the tubes at room temperature for 10 minutes
• Incubate the tubes in a tube rack or lying flat on the lab bench or table.
B. Add DNA sample
Next, follow the steps below to add the DNA Sample to the Cas9/gRNA complex.
6. Add 5 μl of DNA Sample to each of the tubes
• Use a micropipette to add the DNA Sample.
• Remember to change tips at each step!
• The total volume in each tube should be 20 μl after completing this step.
Use a micropipette to add each
of the reagents. Remember to
change tips at each step!
7. Thoroughly mix the reagents by pipetting up and down
10 times, then cap the tubes
• If you have a vortexer, you can mix the reagents by
vortexing for 5 seconds.
8. Make sure all the liquid volume collects at the bottom of
the tube
• If necessary, spin the tubes briefly using a microcentrifuge or
shake down with a flick of the wrist.
9. Incubate the tubes at 37 °C for 15 minutes
• Use a miniPCR
®
in heat block mode, a 37 °C incubator, or
other 37 °C heat source.
DNA
Tube A Tube B Tube C Tube D
DNA Sample
5 μl 5 μl 5 μl 5 μl
TOTAL VOLUME 20 μl 20 μl 20 μl 20 μl
10
min
15
min
