PCR Analysis of Tail DNA

Revision as of 14:43, 28 May 2012 by Davebridges (Talk | contribs) (added categories)

Revision as of 14:43, 28 May 2012 by Davebridges (Talk | contribs) (added categories)

see Genotyping Details for strain specific details

Materials

Protocol

Use the following Volumes per Reaction:

  1. Buffer: 4 uL of 5X Go-Taq buffer ("Molecular Biology Stuff" box in freezer)
  2. Forward Primer: 0.4ul
  3. Reverse Primer: 0.4ul
  4. dNTPs: 0.4uL of 2 mM ("Molecular Biology Stuff" box in freezer)
  5. Sterile water: 13.6 uL
  6. Polymerase Go-Taq: 1 uL (6th floor in Genotype Yellow Box in freezer)
  7. Template: 1 uL

Run PCR Program (approx 2 hours). Use Cycler 1 on 6th Floor

  • Login: Sergey, Just press enter to Login
  • Under Genotype folder, pick Ingles program for Ingles genotyping
  • Under Genotype folder, pick regpcr program for PLT genotyping

Make sure to press enter 2x once to confirm Tubes and second time to start PCR

see Preparing an Agarose Gel for details on preparing a DNA gel