Lab 305: Part I, Day 1
Restricting DNA Samples ("crime scene" or "suspect")
- Each team should obtain 5 reaction tubes. Label them:
- U (DNA from the crime scene, uncut control)
- CS (DNA from the crime scene, will be cut with enzyme)
- 1 (DNA from suspect #1, will be cut with enzyme)
- 2 (DNA from suspect #2, will be cut with enzyme)
- 3 (DNA from suspect #3, will be cut with enzyme)
- Obtain DNA, enzyme and necessary reagents as directed by your teacher. Keep the DNA, buffer,
and enzyme ON ICE. Add the appropriate solutions as indicated below:
- Tube U: 6 µL 2X restriction buffer
2 µL "U" DNA
4 µL sterile H2O
- Tube CS:
6 µL 2X restriction buffer
2 µL "CS" DNA
4 µL enzyme
-
Tube1:
6 µL 2X restriction buffer
2 µL "SUSPECT 1" DNA
4 µL enzyme
-
Tube 2:
6 µL 2X restriction buffer
2 µL "SUSPECT 2" DNA
4 µL enzyme
-
Tube 3:
6 µL 2X restriction buffer
2 µL "SUSPECT 3" DNA
4 µL enzyme
- After all three solutions have been added to each tube, close the caps tightly and mix by giving the tubes a quick 2-3 second spin in a microcentrifuge. (BE SURE TUBES ARE BALANCED IN THE ROTOR -- YOU'LL NEED A SIXTH TUBE AS A BALANCE TUBE.)
- Place your tubes in a 37 degree C water bath for at least 30 minutes. Let stand overnight at room temperature or store in a refrigerator.
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