User:Tara K. Luckau/Notebook/Team ConGen/2010/11/09

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Meeting with Rulon - PCR fixes

  1. try Scun22 primers
    • maybe Scun2 just doesn't cross-amplify between Sceloporus undulatus and Sceloporus occidentalis
  2. check math on dNTPs, ladder (too bright?)
    • maybe I didn't make them correctly
  3. check pH of Tris-Cl and Low TE
    • the current working stocks of both are being stored in 50mL conicals instead of glass
  4. try positive PCR product straight into gel
    • is my ladder too concentrated or is my DNA not amplifying
  5. PCR Rulon's timber rattlers
    • Box "C. horridus Primer stocks"
      • use #9 (needs rehydration)
      • use his Plat Taq, 15mM MgCl2, 10x PCR Buffer, dNTPmix
    • Box "C. horridus NH DNA extract 9-17-8"
      • extracted DNA, but from different locale
      • NanoDrop before using (don't know concentrations)
  6. try better quality Taq
    • may need higher fidelity, or ice wasn't enough