Genotyping protocol

  1. Cut the tail for about 0.5~1cm.
  2. Add 200µl Direct PCR lysis buffer and 10 µl proteinase K (20mg/ml, -20°C).
  3. Incubate at 65°C overnight.
  4. Heat samples at 85°C for 45 min to inactivate proteinase K.
  5. Genotyping PCR.
    (μl)
    5 ×Gotaq PCR Buffer 4
    MgCl2 (50mM) 0.8
    dNTPs (10mM) 0.5
    Primer Mix (10μM) 1
    Tail DNA 2
    Gotaq polymerase 0.2
    ddH20 11.5
    Total 20

    Step 1: 95°C, 4min
    Step 2: 95°C, 30 sec
    Step 3: 55 or 60°C, 30 sec
    Step 4: 72°C, 1 min
    Step 5: Repeat steps 2-4 for 40 cycles
    Step 6: 72°C,10 min.
  6. Run 2% or 3% gel.