Days to Teach:
DNA extraction (20 min), PCR mini-lecture (15 min - can do after setting up PCR to run), Creating Primers / Primer-3 (30 min), PCR (10 min set-up and 1 hr to run), mtDNA comparisons (80 min)
Teaching Tips / Activity Overview:
- DNA Extraction. Use DNA Extraction worksheet.
- PCR lecture
- Creating Primers. Use the Creating Primers worksheet. Do Primer3 activity before handing out PCR procedure worksheet, as the PCR worksheet contains the actual sequence for the primers used.
- PCR. Use the PCR Procedure worksheet.
- Send samples to ColdSpring for sequencing.
Day 3 - mtDNA Comparison. Will be ~ 1 week after PCR procedure to give time for DNA to be sent off and sequences returned.
- Use the mtDNA Comparisons worksheet (see attached documents)
- Use the website to obtain student results.
- Use Dolan site for phylogenetic activity.
- Students will need to make sure they only enter the most reliable portion (that which does not have a lot of discrepancy on bases) of their sequence into the BioServer when doing mtDNA comparison.
For PCR Amplification:
- You can use Science in Motion to borrow necessary equipment if you do not have it readily available in your school.
- You can order materials for the procedure (Carolina and WARDs each sell kits/materials) or you can make your own.