DNA Extractions from 46 Samples of D. innubila Injected with Dilutions of Passage 4 DiNV and Frozen at Days 0, 1, 3, and 5 Post Injection Round 2
The goal is to do qPCR on these samples to track viral load over time, so the DNA needs to be extracted. All samples were thawed on ice, this general protocol was used, and filter tips were used at all times. There were 2 extraction controls used. I ordered the samples so that I started with the earliest day and the lowest dilution and moved up to the higher dilution, then went to the next day to presumably have the lest infected flies at the start of the extraction round and end with the more infected flies. I tried to keep some of every treatment type in each round of extraction incase one round went poorly. It is thought that every fly should have the virus, although to different levels. DNA pellets were resuspended in 25ul of DNA hydration solution. Because there were over 200 flies, DNA extractions were done in 5 rounds.
Samples processed:
| tube | sex | treatment | day | vial |
|---|---|---|---|---|
| ex ctr 3 | NA | NA | NA | NA |
| 13 | female | 0.1 FFU | day0 | NA |
| 14 | female | 0.1 FFU | day0 | NA |
| 23 | male | 0.01 FFU | day0 | NA |
| 24 | male | 0.01 FFU | day0 | NA |
| 31 | female | 0.01 FFU | day0 | NA |
| 32 | female | 0.01 FFU | day0 | NA |
| 39 | male | 3 FFU | day0 | NA |
| 40 | male | 3 FFU | day0 | NA |
| 47 | female | 3 FFU | day0 | NA |
| 48 | female | 3 FFU | day0 | NA |
| 55 | male | 0.01 FFU | day1 | 7 |
| 56 | male | 0.01 FFU | day1 | 7 |
| 65 | female | 0.01 FFU | day1 | 8 |
| 66 | female | 0.01 FFU | day1 | 8 |
| 74 | male | 0.1 FFU | day1 | 1 |
| 75 | male | 0.1 FFU | day1 | 1 |
| 84 | female | 0.1 FFU | day1 | 2 |
| 85 | female | 0.1 FFU | day1 | 2 |
| 93 | male | 3 FFU | day1 | 13 |
| 94 | male | 3 FFU | day1 | 13 |
| 101 | female | 3 FFU | day1 | 14 |
| 102 | female | 3 FFU | day1 | 14 |
| 109 | male | 0.01 FFU | day3 | 9 |
| 110 | male | 0.01 FFU | day3 | 9 |
| 118 | female | 0.01 FFU | day3 | 10 |
| 119 | female | 0.01 FFU | day3 | 10 |
| 128 | male | 0.1 FFU | day3 | 5 |
| 129 | male | 0.1 FFU | day3 | 5 |
| 137 | female | 0.1 FFU | day3 | 6 |
| 138 | female | 0.1 FFU | day3 | 6 |
| 146 | male | 3 FFU | day3 | 15 |
| 147 | male | 3 FFU | day3 | 15 |
| 155 | female | 3 FFU | day3 | 16 |
| 156 | female | 3 FFU | day3 | 16 |
| 163 | male | 0.01 FFU | day5 | 11 |
| 164 | male | 0.01 FFU | day5 | 11 |
| 171 | female | 0.01 FFU | day5 | 12 |
| 172 | female | 0.01 FFU | day5 | 12 |
| 180 | male | 0.1 FFU | day5 | 3 |
| 181 | male | 0.1 FFU | day5 | 3 |
| 191 | female | 0.1 FFU | day5 | 4 |
| 192 | female | 0.1 FFU | day5 | 4 |
| 201 | male | 3 FFU | day5 | 17 |
| 202 | male | 3 FFU | day5 | 17 |
| 209 | female | 3 FFU | day5 | 18 |
| 210 | female | 3 FFU | day5 | 18 |
| ext ctr 4 | NA | NA | NA | NA |