01 - Forensic Escape Room: Design Your Own Biotech Adventure
Module III: DNA Fingerprinting After using blood typing to determine which samples are from the suspect, you must use DNA fingerprinting to find out who the blood came from. You send the suspects’ 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 samples obtained from the suspects and compare them to the blood found at the scene.
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Concentrated buffer
Distilled water
Agarose
60°C
Flask
Caution! Flask will be HOT!
WAIT
POUR
60°C
ADD diluted SYBR Safe
60°C
CASTING THE AGAROSE GEL 1. DILUTE concentrated (50X) buffer with distilled water to create 1X buffer (see Table A).
REMINDER: This kit requires 0.8% agarose gels cast with at least 4 wells.
2. MIX agarose powder with 1X buffer in a 250 mL flask (see 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 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. Before casting the gel, ADD diluted SYBR® Safe to the molten agarose and swirl to mix (see Table A). 7. 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. 8. REMOVE end caps and comb.
Take particular care when re- moving the comb to prevent damage to the wells.
Individual 0.8% UltraSpec-Agarose™ with SYBR ® Stain
7 x 7 cm
0.6 mL
29.4 mL
0.24 g
30 mL
30 µL
10 x 7 cm*
0.9 mL
44.1 mL
0.36 g
45 mL
45 µL
14 x 7 cm
1.2 mL
58.8 mL
0.48 g
60 mL
60 µL
* Recommended gel volume for the EDGE™ Integrated Electrophoresis System.
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