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
- Used thermal cycler in room 215 (because used strips with caps, not plate)
|