In this hands-on lab, students will delve into the history of CRISPR by exploring the inner workings of CRISPR-Cas9 complex in prokaryotes by designing their own synthetic guide RNA (sgRNA) and simulating the action of CRISPR-Cas9 in recognizing and cutting a DNA sequence at its complementary site. Then, students compare and contrast the DNA repair mechanisms in eukaryotes and discover how these can be harnessed, along with CRISPR-Cas9 technology, to precisely knock-out or knock-in genes of interest in eukaryotes. Finally, gel electrophoresis is used to demonstrate the successful CRISPR-Cas9 cleavage or insertion of a target gene sequence.
Materials Included in each MiniLab
Each MiniLab contains enough materials for 10 workstations, 2 – 3 students per workstation.
- Four Ready-to-Load DNA samples
- MiniOne® Universal Marker
- Ten 1% agarose GreenGel™ GelCups
- Two 50 mL BufferCups Tris-Borate-EDTA (TBE) buffer concentrate
- One bag of 0.65 mL microcentrifuge tubes
- One bag of 2 – 200 µL micropipette tips
- 10 sets of activity cards