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./29
Critical thinking
1. To test whether Cas9 will cut where you predicted, you
will run your resulting DNA fragments on an agarose
gel. To the right is a schematic gel image with a DNA
ladder of known DNA sizes. For each of your gRNAs,
draw where you expect the resulting two DNA bands to
be on the gel, using the marked ladder as a guide. As
an example, the uncut DNA sample band (of 3,142 base
pairs) has been done for you.
2. In today's experiment, you will set up four different
reactions, each containing a different combination of
reagents. This will allow you to investigate the function
of each component in the CRISPR/Cas9 system. Based
on what you have learned in the Background information
section, predict what you expect to see in each reaction.
The table below outlines what you will add to each tube.
Use the information from the table above to predict your experimental results.
Tube A Tube B Tube C Tube D
Nuclease
- Cas9 Cas9 Cas9
Guide RNA
- - gRNA1 gRNA2
DNA
DNA Sample DNA Sample DNA Sample DNA Sample
Tube A Tube B Tube C Tube D
Will the DNA be cut
in this reaction?
Justification
Predicted DNA
fragment size(s)
(from question 6 in
the pre-lab activity
on page 19 & 20)
