p47, TPI, and CO1 PCRs on all other MB DiNV infected flies
Flies had been infected here and here, and previously I had just extracted and PCRd the ones alive at day 14 here and here. Now I went back and extracted all individuals here, and now I had to run PCR on them:
Information on these flies can be found here and here
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.
First 96 samples were run at once:
TPI and CO1 master mix
- 500ul GoTaq
- 25ul TPI F
- 25ul TPI R
- 25ul CO1 F
- 25ul CO1 R
- 300ul molecular grade water
p47 master mix
- 500ul GoTaq
- 25ul p47 F
- 25ul 47 R
- 350ul molecular grade water
Then a second set of 24:
TPI and CO1 master mix
- 132.5ul GoTaq
- 6.625ul TPI F
- 6.625ul TPI R
- 6.625ul CO1 F
- 6.625ul CO1 R
- 79.5ul molecular grade water
p47 master mix
- 132.5ul GoTaq
- 6.625ul p47 F
- 6.625ul 47 R
- 92.75ul molecular grade water
After the PCRs, gels were run for 35 minutes at 90V, and then band presence was scored:
| tube | treatment | day | dead | TPI | CO1 | p47 |
|---|---|---|---|---|---|---|
| 3 | 3 FFU | day3 | yes | yes | yes | yes |
| 5 | 0.01 FFU | day6 | yes | yes | yes | no |
| 6 | 0.05 FFU | day6 | yes | yes | yes | yes |
| 7 | 0.05 FFU | day6 | yes | yes | yes | yes |
| 8 | 0.05 FFU | day6 | yes | yes | yes | yes |
| 9 | 0.05 FFU | day6 | yes | no | no | yes |
| 10 | 3 FFU | day6 | yes | yes | yes | yes |
| 11 | 3 FFU | day6 | yes | yes | yes | yes |
| 13 | 3 FFU | day6 | yes | yes | yes | yes |
| 14 | CCM | day9 | yes | yes | no | yes |
| 15 | 0.01 FFU | day9 | yes | yes | yes | yes |
| 16 | 0.01 FFU | day9 | yes | yes | yes | yes |
| 17 | 0.01 FFU | day9 | yes | yes | yes | yes |
| 18 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 19 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 20 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 21 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 22 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 23 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 24 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 26 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 27 | 1 FFU | day9 | yes | yes | yes | yes |
| 28 | 1 FFU | day9 | yes | yes | yes | yes |
| 29 | 1 FFU | day9 | yes | yes | yes | yes |
| 31 | 1 FFU | day9 | yes | yes | yes | yes |
| 32 | 1 FFU | day9 | yes | yes | yes | yes |
| 33 | 1 FFU | day9 | yes | yes | yes | yes |
| 34 | 1 FFU | day9 | yes | yes | yes | yes |
| 35 | 1 FFU | day9 | yes | yes | yes | yes |
| 36 | 1 FFU | day9 | yes | yes | yes | yes |
| 37 | 1 FFU | day9 | yes | yes | yes | yes |
| 38 | 1 FFU | day9 | yes | yes | yes | yes |
| 39 | 1 FFU | day9 | yes | yes | no | yes |
| 40 | 1 FFU | day9 | yes | yes | yes | yes |
| 41 | 1 FFU | day9 | yes | yes | yes | yes |
| 42 | 1 FFU | day9 | yes | yes | no | yes |
| 43 | 3 FFU | day9 | yes | yes | yes | yes |
| 44 | 3 FFU | day9 | yes | yes | yes | yes |
| 45 | 3 FFU | day9 | yes | yes | yes | yes |
| 46 | 3 FFU | day9 | yes | no | no | yes |
| 47 | 3 FFU | day9 | yes | yes | yes | yes |
| 48 | 3 FFU | day9 | yes | yes | yes | yes |
| 50 | 3 FFU | day9 | yes | yes | yes | yes |
| 51 | 3 FFU | day9 | yes | yes | yes | yes |
| 52 | 3 FFU | day9 | yes | yes | yes | yes |
| 1 | 0.05 FFU | day3 | yes | yes | yes | no |
| 3 | 0.05 FFU | day6 | yes | yes | yes | yes |
| 4 | 0.05 FFU | day6 | yes | yes | yes | yes |
| 5 | 0.05 FFU | day6 | yes | yes | yes | yes |
| 6 | 1 FFU | day6 | yes | yes | yes | yes |
| 8 | 1 FFU | day6 | yes | yes | yes | yes |
| 9 | 3 FFU | day6 | yes | yes | yes | yes |
| 10 | 3 FFU | day6 | yes | yes | yes | yes |
| 11 | 3 FFU | day6 | yes | yes | yes | yes |
| 12 | 0.01 FFU | day9 | yes | yes | yes | no |
| 13 | 0.01 FFU | day9 | yes | yes | no | yes |
| 14 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 15 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 16 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 17 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 18 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 19 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 20 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 21 | 0.05 FFU | day9 | yes | yes | yes | yes |
| 22 | 0.05 FFU | day9 | yes | yes | no | yes |
| 23 | 0.05 FFU | day9 | yes | yes | no | yes |
| 24 | 1 FFU | day9 | yes | yes | yes | yes |
| 25 | 1 FFU | day9 | yes | yes | yes | yes |
| 26 | 1 FFU | day9 | yes | yes | yes | yes |
| 27 | 1 FFU | day9 | yes | yes | yes | yes |
| 28 | 1 FFU | day9 | yes | yes | no | yes |
| 29 | 1 FFU | day9 | yes | yes | yes | yes |
| 30 | 1 FFU | day9 | yes | yes | yes | yes |
| 31 | 1 FFU | day9 | yes | yes | no | yes |
| 32 | 1 FFU | day9 | yes | yes | yes | yes |
| 33 | 1 FFU | day9 | yes | yes | yes | yes |
| 34 | 1 FFU | day9 | yes | yes | yes | yes |
| 35 | 3 FFU | day9 | yes | yes | no | yes |
| 36 | 3 FFU | day9 | yes | yes | yes | yes |
| 37 | 3 FFU | day9 | yes | yes | yes | yes |
| 38 | 3 FFU | day9 | yes | yes | yes | yes |
| 39 | 3 FFU | day9 | yes | yes | yes | yes |
| 40 | 3 FFU | day9 | yes | yes | yes | yes |
| 41 | 3 FFU | day9 | yes | yes | yes | yes |
| 42 | 3 FFU | day9 | yes | yes | no | yes |
| 43 | 3 FFU | day9 | yes | yes | yes | yes |
| 44 | 3 FFU | day9 | yes | yes | yes | yes |
| 45 | 3 FFU | day9 | yes | yes | yes | yes |
| 46 | 3 FFU | day9 | yes | yes | yes | yes |
| 47 | 3 FFU | day9 | yes | yes | yes | yes |
| 48 | 3 FFU | day9 | yes | yes | yes | yes |
| 49 | 3 FFU | day9 | yes | yes | yes | yes |
| 53 | 3 FFU | day9 | yes | yes | yes | yes |
| 54 | 3 FFU | day9 | yes | yes | yes | yes |
| 60 | 1 FFU | day12 | yes | yes | yes | yes |
| 61 | 1 FFU | day12 | yes | yes | no | yes |
| 62 | 1 FFU | day12 | yes | yes | yes | yes |
| 63 | 1 FFU | day12 | yes | yes | yes | yes |
| 64 | 3 FFU | day12 | yes | yes | yes | yes |
| 65 | 3 FFU | day12 | yes | yes | no | yes |
| 53 | 1 FFU | day12 | yes | yes | yes | yes |
| 54 | 1 FFU | day12 | yes | yes | yes | yes |
| 55 | 1 FFU | day12 | yes | yes | yes | yes |
| 56 | 1 FFU | day12 | yes | yes | yes | yes |
| 57 | 1 FFU | day12 | yes | yes | yes | yes |
| 58 | 1 FFU | day12 | yes | yes | yes | yes |
| 59 | 3 FFU | day12 | yes | yes | yes | yes |
| 60 | 3 FFU | day12 | yes | yes | yes | yes |