PCR Teacher Manual
Materials Included in Kit
This experiment is designed for 15 lab groups of 2 students per each group.
Note: Each group is sharing some materials but each student is performing their own experiment.
Materials Checklist
 1 Teacher’s guide
 1 Student’s guide
 15 tubes
Lysis Buffer, containing 1100 l each
 60
Sterile Microcentrifuge Tubes
 30
Sterile Cotton Swabs
 30 tubes
10% Chelex in sterile water, containing 100 l each
 90
PCR tubes containing a Ready-To-Go PCR Bead
 15 tubes
Sterile Mineral Oil, containing 200 l each
 15 tubes
Sterile Water, containing 75 l each
 10 grams
Agarose Powder
 1500 mL
5X TBE Buffer
 Methylene Blue DNA Stain
 Staining Dishes
Redemption coupon for the following materials:
 30 tubes
Primer A, containing 40 l
 30 tubes
Primer B, containing 40 l
 Control human DNA Allele Samples
o
30 tubes Allele #1 (with Alu insertion-400 bp), containing 7 l
o
30 tubes Allele #2 (without Alu insetion-100 bp), containing 7 l
 15 tubes
Loading Dye, containing 75 l
 15 tubes
DNA Marker Standard (Hae III Digest of X174), containing 25 l
SPECIAL HANDLING INSTRUCTIONS
 Redeem coupon for perishable materials. Note: Should send within 4-6 weeks to
be received on time. Upon receipt place the materials in the freezer.
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MATERIALS NEEDED BUT NOT INCLUDED IN KIT
p20, p2000, p1000 Micropipettes
p20/p200 Micropipette Tips
p1000 Micropipette Tips
Waste container
Marking Pens
Microcentrifuge
Liquid Waste Container
Boiling Water Bath
Floating Rack
Timers (optional)
Ice
Thermocycler
30 Test tubes for making agarose gel
Containers to hold test tubes
 Spatula
 Electronic Balances
 15, 10mL disposable
pipettes
 15, 10 mL pipette pumps
 Vortex (optional)
 Electrophoresis Setup
 Casting deck and casting
tray
 Comb
 Electrophoresis box
 Wires
 Power Supply
Objective:
Students will use Alu PCR amplification and agarose gel electrophoresis techniques to
determine their own Alu genotypes* and genotypes* of control samples.
*The Alu insertion is not a gene. However it simulates the inheritance pattern one would
associate with a gene.
Time Requirements:

 Day 1: DNA Isolation: 45 minutes
Day 2-3: DNA Amplification: 90 minutes
Pre-Lab Preparation
The following dilution can be prepared several days in advance.
 Prepare 1X TBE Buffer: Add 5850 mL distilled water to 650 mL of 10X TBE
concentrate to obtain 6500 mL of 1X buffer.
Day of Lab Experiment (Part 1):
 Set-up boiling water bath (Note: Allow ample time for the water to reach boiling)
Day of Lab Experiment (Part 2):
 Ice for reagents
Lab Station Set Up
Set up for groups of 2 students per station
Part 1:
p20, p200, p1000 Micropipettes
p20/p200 Micropipette Tips
p1000 Micropipette Tips
Marking Pens
Lysis Buffer, 1100 l
4, Sterile Microcentrifuge Tubes
2, Sterile Cotton Swabs
2 tubes containing 100 l of
Chelex
Liquid Waste Container
Solid Waste Container
Ice
Equipment:
Microcentrifuges
Boiling Water Bath w/ chips
Floating Rack
Timers (optional)
Part 2:
p20, p200, Micropipettes
p20/p200 Micropipette Tips
Marking Pens
Solid Waste Container
6, PCR tubes containing a
Ready- To-Go PCR Bead
Sterile Mineral Oil, 150 l
Sterile Water, 40 l
On Ice:
Primer A, 60+ l
Primer B, 60+ l
Student DNA from Part 1
Control DNA Allele Samples
Allele #1 (400 bp), 5+ l
Allele #2 (100 bp), 5+ l
Equipment:
Thermocycler
Part 3:
Test tubes for making gel
Containers to hold test tubes
Spatula
10mL disposable pipettes
10 mL pipette pumps
1X TBE Buffer (500 + mL)
Electrophoresis Setup:
Casting deck & casting tray
Comb
Electrophoresis box
Wires
Power Supply
On Ice:
Loading Dye, 60+ l
DNA Marker Standard ,
20+ l
Weighing stations
Agarose Powder
Electronic Balances
Vortex (optional)
Staining
DNA Stain
Staining Dishes
UV box, for SAFE STAIN