After yesterday’s confirmation that the primer/probe sets for RLOv DNA helicase and head-to-tail were functional and specific for the RLOv (and don’t amplify RLO alone), I needed to confirm that the qPCRs only generated a single product in each reaction via melt curve analysis.
NOTE: Remaining volume of template DNA wasn’t going to be sufficient for all reactions, so added 100μL of NanoPure H2O. Seeing how early the amplification was in yesterday’s qPCR (Cq ~15), this dilution should be fine.
All samples were run in duplicate.
Master mix calcs are here: 20151009 – qPCR RLOv
Plate layout, cycling params, etc can be found in the qPCR Report (see Results).
Both primer sets amplified a single PCR product. This is demonstrated by the single melt peak for each primer set.