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Lab 305: Part I, Day 1
Restricting DNA Samples ("crime scene" or "suspect")

  1. 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)

  2. 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


  3. 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.)

  4. 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.


Background     Part II & III     Part IV     Part V     Part VI

Crime Scenario     Historical Mystery     Teacher's Guide




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