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

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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)


 * [[Image:2011 03 11 PCR.jpg|900 px]]

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
 * [[Image:20110311_GelFull.jpg|800 px]]


 * Zoom in on the only part that had amplicon:
 * [[Image:20110311_GelZoom.jpg|800 px]]
 * 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


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