1. BIO-RAD Forensic DNA Fingerprinting Kit
Section Concepts: DNA Scissors Activity, restriction enzymes, gel electrophoresis, standard curve

2. Background Information
Restriction enzymes: found in bacteria and archaea, are enzymes that cuts DNA at specific recognition nucleotide sequences known as restriction sites.
Gel Electrophoresis: a method used in biochemistry and molecular biology to separate a mixed population of DNA and RNA fragments by length, to estimate the size of DNA and RNA fragments, or to separate proteins by charge. Nucleic acid molecules are separated by applying an electric field to move the negatively charged molecules through an agarose matrix.

3. Restriction Enzymes + Gel Electrophoresis
Restriction enzymes video
Gel Electrophoresis animation

4. Timeline/Overview Monday: Lecture
Tuesday: Activities 1 & 2, DNA scissors, pre-lab quiz
Thursday: DNA fingerprint (restriction digest), Analyze lab (work on packets)
Monday: No school
Tuesday: DNA fingerprinting kit (run gel, visualize)
Thursday: Packet due, post quiz

5. Necessary Preparation for Lab DNA Finger printing
Labels
8—clear 1.5mL tubes ENZ (enzyme)
8—clear 1.5mL tubes M (marker)
8—clear 1.5mL tubes LD (loading dye)
8--green 1.5mL tubes CS
8—blue 1.5mL tubes S1
8—orange1.5mL tubes S2
8—violet 1.5mL tubes S3
8—red 1.5mL tubes S4
8—yellow 1.5mL tubes S5
Before Digest
Set water bath for 37C
Rehydrate & aliquot (page 13)
EcoRI/PstI enzyme mix (12 hours before use)
Crime scene DNA
Suspects 1-5 DNA
Before Running Gel
Prepare electrophoresis TAE buffer
11/13 at before 5th period Pour 8 gels (1%, EtBr, 10 well comb)
Prep. HindIII standard (right before class)
Aliquot
HindIII (marker)
Loading dye

6. DNA Fingerprinting Lab - Background Information
DNA Fingerprinting: technique used by scientists to identify an individual based on their specific DNA profile
Restriction Digestion of DNA: restriction enzymes recognize particular sequences in a segment of DNA and cuts the DNA molecule at that point
Plasmids: circular, non-chromosomal pieces of DNA that can replicate in and are commonly found in bacteria

7. DNA Fingerprinting
DNA Fingerprinting Video

8. Plasmids Restriction Enzyme Digestion of Plasmid DNA
Restriction Enzyme Animation
Restriction Enzyme EcoR1 Animation

9. DNA Fingerprinting Tips
Place tubes in refrigerator after removing from water bath
Gel will be 10 wells, adjust lanes in your protocol to use 2-8