From 2007.igem.org

• about PCR issue for INS_A and INS_B
  • so far we have two PCR (9/5 and 9/8)
  • Because the concentration of 9/8 is too low, we suspect that the temperature could cause the problem
  • five annealing temperatures are tested in INS_A
    • #1: 49.8
    • #2: 52.3
    • #3: 55.5
    • #c: 55.5 (control without template)
    • #4: 58.7
    • #5: 60.9
    • PCR protocol
      • template 1.0 uL
      • 10X PCR buffer 5.0 uL
      • dNTP (2.5mM), 2.0 uL
      • primer F 1.0 uL
      • primer R 1.0 uL
      • polymerase 0.5 uL
      • ddH2O 39.5 uL
      • total is 50.0 uL