User:Tara K. Luckau/Notebook/Team ConGen/2011/03/18

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Scun2 FAM Diversity Panel PCR

  • chose Buffer G, 58°C as 'best' condition for Scun2 primers (with FAM)
  • now run diversity panel and submit to fragment analysis

Diversity Panel

Sceloporus occidentalis from all five sites (plus two redundancies)

  • SCOC RAJ X01 - Rancho Jamul
  • SCOC HOL X01 - Hollenbeck
  • SCOC SYR2 X01 - Santa Ysabel
  • SCOC CPN X01 - Camp Pendleton
  • SCOC TP1 X01 - Torrey Pines
  • SCOC TP3 X01 - Torrey Pines
  • SCOC LOM X01 - Point Loma


PCR

  • First PCR to send to fragment analysis!
  • use labelled forward primer (Scun2 F FAM)
  • increase reaction volume (10µL to 15µL) to have enough to ship to Tucson after confirmation gel
  • gel results from gradient PCR was a bit weird (only Buffer G yielded any product), so use three buffers: F, G, H
  • this also makes shipping to Tucson worth the money
  • 20110318 PCR.jpg
  • Used thermal cycler in room 215 (because used strips with caps, not plate)