DNA Extractions and PCRs on Stock D. innubila Genome to Check for DiNV Infection
PCR data from the freezing experiment showed a lot of contamination of DiNV in the sterile pricked flies. This could be because my stock flies are infected some how. I wanted to check my flies to see if there was any evidence of DiNV in them. I took 6 male flies that had been separated with CO2, but not had been on the CO2 pad. I separated them on a petri dish with the CO2 flowing on it, and the fly room utensils were used. The other 6 flies were of unknown sex and were just frozen in their vial, never were on CO2 and never touched a fly room utensils.
DNA extractions from these samples followed exactly the single fly DNA extraction protocol.
Sample information can be found here
PCRs on CO1, TPI, and p47 were done accordingly to the general PCR protocol with the primers, and their information can be found here. The p47 samples were amplified in 30 cycles.
A 2% gel was run for 45 minutes at 90V to analyze the PCRs.
All samples are DiNV negative, so this is looking good for my stock flies not being infected. However these flies were all the same age, about 0-3 days old. It could be that older flies show the infections. I will need to check more to be sure.