Introduce your students to the concepts of symbiotic relationships, DNA extraction, PCR, and electrophoresis techniques with the insects provided in the kit. Using the insects and a positive and negative control, students will be able to test for the presence of Wolbachia, an intracellular bacterium found in approximately 20% of all insect species. The insect DNA will be amplified using specific primers to detect a sequence that is unique to Wolbachia, allowing students to easily see whether their mystery insects have this bacteria. Once you have completed the lab, you may run the experiment again with insects that you collect on your own to see how they compare. The activity comes with enough materials to run 24 PCR reactions, a teacher’s guide, and student copymaster. Note: Coupon included for perishable materials. Redeem by mail, fax, phone, or e-mail. Ethidium bromide version sold only to colleges, universities, and accredited laboratories. Check your local regulations for disposal of ethidium bromide.
Materials Included
•1 Bottle, cell lysis solution,
•1 Bottle, protein precipitation solution,
•1 Bottle, 99% isopropyl alcohol,
•1 Bottle, 70% ethanol,
•1 Bottle, DNA hydration solution,
•1 Vial, DNA dilution buffer,
•30 Microfuge tubes,
•1 Vial, sterile mineral oil,
•24 PCR Reaction tubes,
•6 Cotton swabs, sterile,
•1 Bottle, 2.0% agarose,
•1 Bottle, TBE buffer, 5X,
•1 Bottle, DNA stain,
•1 Vial, Loading dye,
•1 Vial, Primer W-spec forward,
•1 Vial, Primer W-spec reverse,
•1 Redemption coupon for: 2 Vials, ?X174/HaeIII DNA, 1 Vial, weight marker, 1 Bottle, RNAse, 1 Bottle, proteinase K, 1 Vial, PCR master mix, 1 Vial, Nasonia positive control, 1 Vial, Nasonia negative control, 1 Vial, mixed insects (unknowns)
Materials Needed But Not Provided
•Thermal cycler for PCR
•Hot water bath or incubator at 65°C
•Hot water bath or incubator at 37°C
•Ice bath
•Micropipet
•Micropipet tips
•Electrophoresis tank apparatus and power supply
•Paper towels
•Staining tray
•Microcentrifuge
Time Requirement
•Part I: 45 minutes
•Part II: 20 minutes setup/90 minutes to run PCR
•Part III: 90 minutes
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