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

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==Task==
==Scun2 FAM Gradient PCR==
* here
* Moving on with Scun2:
* here
:* use labelled forward primer (Scun2 F FAM) (get ready to genotype)
** in here
:* full temperature and MgCl<sub>2</sub> gradient to find annealing temperature (no touchdown)
# numbered
* [[Image:2011 03 11 PCR.jpg|900 px]]
# again


==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]]


===Subsection===
 
* here
* 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




Revision as of 15:16, 18 March 2011

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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



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