CO1, TPI, and p47 PCR of Flies Alive on Day 14 MB Infections to Check for Virus Presence
Checking flies at the end of the experiment for presence of DNA to see if it looks like 0.01 FFU does flies received any virus
Information on the primers and their programs can be found here
The PCR process followed this general protocol. CO1 and TPI were run in the same reaction.
TPI and CO1 master mix
- 260ul GoTaq
- 13ul TPI F
- 13ul TPI R
- 13ul CO1 F
- 13ul CO1 R
- 156ul molecular grade water
p47 master mix
- 260ul GoTaq
- 13ul p47 F
- 13ul 47 R
- 182ul molecular grade water
After the PCRs, a gel was run for 35 minutes at 90V, and then band presence was scored:
| tube | treatment | day | day | dead | extracted | TPI | CO1 | p47 |
|---|---|---|---|---|---|---|---|---|
| ex ctrl 1 | NA | NA | NA | NA | 20240329 | no | no | no |
| 81 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 82 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 83 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 84 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | yes |
| 85 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 86 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 87 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 88 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 89 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 90 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 91 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 92 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 93 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 94 | 0.01 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 95 | 0.05 FFU | day14 | 3/5 | no | 20240329 | yes | yes | yes |
| 96 | 0.05 FFU | day14 | 3/5 | no | 20240329 | yes | yes | yes |
| 97 | 0.05 FFU | day14 | 3/5 | no | 20240329 | yes | yes | no |
| 85 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | no |
| 86 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | no |
| 87 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | no |
| 88 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | no |
| 89 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | no |
| 90 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | no |
| 91 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | no |
| 92 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | no | yes |
| 93 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | yes |
| 94 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | yes |
| 95 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | no |
| 96 | 0.01 FFU | day14 | 3/2 | no | 20240329 | yes | yes | yes |
| 97 | 0.05 FFU | day14 | 3/2 | no | 20240329 | yes | yes | yes |
| 98 | 0.05 FFU | day14 | 3/2 | no | 20240329 | yes | yes | yes |
| 98 | 0.01 FFU | day14 | 3/5 | yes | 20240329 | no | no | no |
| 2 | 0.01 FFU | day6 | 3/5 | yes | 20240329 | yes | yes | yes |
| 50 | 0.01 FFU | day12 | 3/5 | yes | 20240329 | yes | no | yes |
| 51 | 0.05 FFU | day12 | 3/5 | yes | 20240329 | yes | yes | yes |
| 52 | 0.05 FFU | day12 | 3/5 | yes | 20240329 | yes | yes | yes |
| 66 | 0.01 FFU | day14 | 3/2 | yes | 20240329 | yes | yes | yes |
| 67 | 0.05 FFU | day14 | 3/2 | yes | 20240329 | yes | yes | yes |
| 55 | 0.01 FFU | day12 | 3/2 | yes | 20240329 | yes | yes | yes |
| 56 | 0.05 FFU | day12 | 3/2 | yes | 20240329 | yes | yes | yes |
| 57 | 0.05 FFU | day12 | 3/2 | yes | 20240329 | maybe | no | yes |
| 58 | 0.05 FFU | day12 | 3/2 | yes | 20240329 | maybe | no | yes |
| 59 | 0.05 FFU | day12 | 3/2 | yes | 20240329 | no | no | yes |
| 4 | 0.01 FFU | day6 | 3/2 | yes | 20240329 | maybe | no | no |
| ex ctrl 2 | NA | NA | NA | NA | 20240329 | no | no | no |