Carolyn & Stan Langevin have agreed that the DNA helicase qPCR should be tested on 10 black abalone DNA extractions that fall into multiple levels of the Friedman Lab’s withering syndrome histology scoring.
Earlier today I identified samples at three different histology scoring levels of RLO: 0, 1, & 2.
Here’s the list of samples that will be qPCR’d. There were only eight samples that had histology scores of 2 in both PE and Dg.
Ran qPCR using the RLOv DNA helicase standard curve from 20151106.
All samples were run in duplicate on the CFX96 (BioRad).
Master mix calcs are here: 20151120 – qPCR RLOv Black Abs
Plate layout, cycling params, etc. can be seen in the qPCR Report (see Results).
qPCR Report (PDF): Sam_2015-11-20 15-00-27_CC009827.pdf
qPCR Data File (CFX96): Sam_2015-11-20 15-00-27_CC009827.pcrd
Quick summary of the results:
- 50% of the RLO/RLOv 0 score samples are positive for RLOv DNA helicase. Will talk to Carolyn to see if she has withering syndrome qPCR data for these samples to compare RLOv-positive samples with WSN-positive samples. If not, will run withering syndrome qPCR.
- All RLO/RLOv 1 & 2 scored samples are positive for RLOv DNA helicase
- All RLO/RLOv 2 scored samples come up before the standar curve; these should be diluted and re-run.
- Standard curve isn’t perfect (the 3 copy sample is throwing it off).
STANDARD CURVE AMP & SCATTER PLOTS
RLO/RLOv 0 AMP PLOTS
RLO/RLOv 1 AMP PLOTS
RLO/RLOv 2 AMP PLOTS