Though not shown in the work-flow, ReadqPCR also allows the user to deal with technical replicates by (optionally) calculating the arithmetic mean or median of the raw Cq values for the technical replicates of a given gene.
Besides these 2 candidates, quite similar average expression levels for the remaining 11 genes, with raw Cq values around 25, were observed.
And we did not mean raw fish.
I mean raw, raw, raw!" The Afghan gave him a quizzical look, as if uncertain whether he was being chastised.
Raw Cq values were exported for further analysis.
As suggested in previous work, genes that showed a coefficient of variance of the raw Cq values greater than 4% were excluded from further evaluation [39].
Third applet uses raw Cq data and determines the most stably expressed genes based on coefficient of correlation (r) to the BestKeeper Index (BI).
All raw Cq values >32 were discarded.
Statistical analysis was performed using GraphPad Prism 6 [ 123] after normalization of raw Cq values.
The raw Cq values obtained for COI were transformed to relative normalized quantities using the Cq values for histone and tubulin by the software qBase.
