DNA Extractions from 33 Samples of D. innubila Injected with Dilutions of Passage 4 DiNV and Frozen at Days 0, 1, 3, and 5 Post Injection Round 5

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 9 NA NA NA NA
5 male 0.1 FFU day0 NA
6 male 0.1 FFU day0 NA
7 male 0.1 FFU day0 NA
8 male 0.1 FFU day0 NA
9 male 0.1 FFU day0 NA
10 male 0.1 FFU day0 NA
19 female 0.1 FFU day0 NA
20 female 0.1 FFU day0 NA
61 male 0.01 FFU day1 7
62 male 0.01 FFU day1 7
71 female 0.01 FFU day1 8
80 male 0.1 FFU day1 1
81 male 0.1 FFU day1 1
90 female 0.1 FFU day1 2
115 male 0.01 FFU day3 9
124 female 0.01 FFU day3 10
125 female 0.01 FFU day3 10
134 male 0.1 FFU day3 5
143 female 0.1 FFU day3 6
152 male 3 FFU day3 15
177 female 0.01 FFU day5 12
186 male 0.1 FFU day5 3
187 male 0.1 FFU day5 3
197 female 0.1 FFU day5 4
198 female 0.1 FFU day5 4
207 male 3 FFU day5 17
208 male 3 FFU day5 17
213 female 3 FFU day5 18
214 female 3 FFU day5 18
215 female 3 FFU day5 18
216 female 3 FFU day5 18
217 female 3 FFU day5 18
218 female 3 FFU day5 18
ex ctr 10 NA NA NA NA