User:Tara K. Luckau/Notebook/Team ConGen/2011/03/11

Tara's Lab Notebook

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Scun2 FAM Gradient PCR Moving on with Scun2: use labelled forward primer (Scun2 F FAM) (get ready to genotype) full temperature and MgCl2 gradient to find annealing temperature (no touchdown) Scun2 FAM Gradient Gel Pour: 270mL 1x TAE + 5.4g agarose + 27µL GelRed Load: 1µL gel load buffer + 5µL PCR product Run: 1 hour, 150V Zoom in on the only part that had amplicon: Amplicon at ~150bp consistent with expected amplicon size - nice and clean amplicon present across many temperatures, but more so on lower range (55-60°C) try: Buffer G, 58°C, 15µL reaction on diversity panel to send to frag