DNA Extraction and TPI/p47 PCRs of A Few Samples from Replicate 2 of the Male and Female D. innubila 16 Cq Injections
DNA Extractions 20231109
I want to make sure that there was no contamination of my controls during this experiment (Taiye had some contamination in his flies and I want to make sure that this was not a result of contaminated materials that we shared). Additionally I would like to do a little more qPCR on flies who died from virus infection (we think), so I need their DNA extracted.
These were the samples chosen for DNA extraction:
| tube | sex | day frozen | treatment |
|---|---|---|---|
| 1 | female | day0 | CCM |
| 2 | female | day0 | CCM |
| 8 | male | day0 | CCM |
| 77 | female | day18 | CCM |
| 78 | male | day18 | CCM |
| 79 | male | day18 | CCM |
| 22 | female | day6 | CCM |
| 74 | female | day18 | CCM |
| 14 | female | day0 | 16Cq DiNV |
| 15 | female | day0 | 16Cq DiNV |
| 17 | male | day0 | 16Cq DiNV |
| 21 | male | day3 | 16Cq DiNV |
| 88 | female | day18 | 16Cq DiNV |
| 46 | female | day9 | 16Cq DiNV |
| 47 | male | day9 | 16Cq DiNV |
| 57 | female | day9 | 16Cq DiNV |
| 62 | male | day10 | 16Cq DiNV |
| 65 | female | day12 | 16Cq DiNV |
You can see all sample information here
DNA extraction followed this protocol exactly. Care was taken to make sure control samples were done in first order, than the infected samples. Two extraction controls were also included in this extraction.
PCRs 20231110
PCRs for TPI and p47 followed this general protocol, where I PCRd every sample including the extraction controls. A negative and positive control for each primer was run. Primer information can be found here
Because of gel constraints, samples were run over 2 separate gels
TPI for all samples but the extraction controls :
p47 for almost all samples:
And the last section on this gel has the 2 extraction controls with TPI, and the remaining p47 samples:
Full information on the samples and their PCR results can be found here
Notably, the one female fly who did not die but got injected with DiNV did not show the virus when the experiment ended.