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

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