User:Tara K. Luckau/Notebook/Team ConGen/2011/01/11

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Contents


Gel Touchdown

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  • Faint bands at very bottom, near next rows' lanes = primer front
    • ladder triplet: 396/344/298
    • ladder doublet: 220/201
    • ladder doublet: 154/134
    • ladder band: 75
  • but no amplicon at expected 450bp

Gel Timber Rattlesnakes

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  • ---


  • Definitely bands! Too bad I ran off primer 10's bands (but I assume they're there)


Next Steps

  • Try to break the PCR by using Tara's dNTPs, Tara's Buffer
  • Try to make Scun PCRs work by using Rulon's dNTPs, 10x Buffer + MgCl2


Extract Sceloporus undulatus

  • acquired from Tod Reeder; "H0074; TWR 78" (looks like muscle and liver tissue)
  • extract same species as primers were designed for, to rule out DNA extraction and primer synthesis as PCR problem
  1. Add 300µL of Cell Lysis Solution to about 10mg tissue
  2. Add 2µL Proteinase K (20mg/mL); incubate overnight at 55°C water bath



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