01 - Forensic Escape Room: Design Your Own Biotech Adventure
Module III: DNA Fingerprinting
After determining the blood collected from the knife is Type A you must use DNA fingerprinting to find out which suspect the blood came from. You send both Sarah Anne and Laura’s blood samples as well as the blood collected from the knife to an external DNA lab. Here they will extract DNA from each sample and amplify it by PCR. In this module, you will use gel electrophoresis to analyze the PCR reactions obtained from the suspects and compare them to the blood found at the scene.
Concentrated buffer
Distilled water
Agarose
60°C
Flask
Caution! Flask will be HOT!
WAIT
POUR
60°C
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 completely dis- solved (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 transparent as it solidifies. 7.
REMOVE end caps and comb. Take particular care when removing the comb to prevent 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
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