p47-q PCR on DiNV Primary Day 15 3-23 Used in Infecting Myd88 and W1118 Flies

I had previously used the p47_q PCR primers on the DNA extracted from the infected flies, but I did not try to amplify the virus in the actual infection liquid. So I did a separate PCR for that.

  • Samples were the infection liquid, and I also did a negative control of water
  • Reaction was made on ice
  • Made master mix on ice:
    • 11ul GoTaq
    • 0.55ul p47_q F
    • 0.55ul p47_q R
    • 7.7ul molec grade water
  • Vortexed and spun down mix
  • Added 9ul of mix to 2 strip tubes on ice
  • Added 1ul of DiNV Primary Day 15 3-23 liquid to the first tube
  • Added 1ul molec grade water to the second tube
  • Vortexed and spun down tubes
  • Placed tubes in the 55TEST program (55 degree anealing temp and 30 sec extension)
  • Samples were placed in the fridge overnight

Gel 20220525

  • 2% gel ran at 100V for ~35 minutes
  • And stained for 35 minutes

Amplification at ~115bp is correct. This is positive for DiNV.