06 - Introducing Your Students to CRISPR with Sickle Cell Gene Editing
Module II: Agarose Gel Electrophoresis
Concentrated buffer
Distilled water
Agarose
60°C
Flask
Caution! Flask will be HOT!
WAIT
POUR
60°C
CASTING THE AGAROSE GEL
1. DILUTE concentrated 50X Electrophoresis buffer with distilled water (refer to Table A for correct volumes depending on the size of your gel casting tray). 2. MIX agarose powder with buffer solution in a 250 mL flask (refer to Table A). 3. DISSOLVE agarose powder by boiling the solution. MICROWAVE the solution on high for 1 minute. Carefully REMOVE the flask from the microwave and MIX by swirling the flask. Continue to HEAT the solution in 15-second bursts until the agarose is com- pletely dissolved (the solution should be clear like water). 4. COOL agarose to 60 °C with careful swirling to promote even dissipation of heat. 5. While agarose is cooling, SEAL the ends of the gel-casting tray with the rubber end caps. PLACE the well template (comb) in the appropriate notch. 6. POUR the cooled agarose solution into the prepared gel-casting tray. The gel should thoroughly solidify within 20 minutes. The gel will stiffen and become less transpar - ent as it solidifies. 7. REMOVE end caps and comb. Take particular care when removing the comb to pre- vent damage to the wells.
Individual 0.8% UltraSpec-Agarose™ Gels
7 x 7 cm
0.6 mL
29.4 mL
0.24 g
30 mL
10 x 7 cm*
0.9 mL
44.1 mL
0.36 g
45 mL
* Recommended gel volume for the EDGE™ Integrated Electrophoresis System. 60 mL *Recommended gel volume for the EDGE™ Integrated Electrophoresis System. (Cat. #500). 14 x 7 cm 1.2 mL 58.8 mL 0.48 g
72
Visit our website at www.edvotek.com
Made with FlippingBook flipbook maker