Reference Gene Stability
Why stability matters
Choosing an unstable reference gene invalidates the entire DDCt analysis. MIQE guidelines recommend validating reference gene stability before use. AnnealIQ provides two independent stability algorithms.
GeNorm M-values
GeNorm ranks reference genes by pairwise variation (M-value). Lower M-values indicate higher stability. Results are shown as a horizontal bar chart with color-coded thresholds: green (highly stable, M < 0.5), yellow (acceptable, M < 1.0), red (unstable, M ≥ 1.0).
GeNorm V-values
GeNorm V-values (pairwise variation) help determine the optimal number of reference genes. A horizontal bar chart shows V-values with a 0.15 threshold line. A plain-English recommendation tells you whether adding another reference gene meaningfully improves normalization.
NormFinder
NormFinder provides an independent stability ranking that accounts for inter- and intra-group variation. Results are displayed alongside GeNorm for comparison.
AI recommendation
After stability analysis, a prominent AI recommendation card summarizes which reference gene(s) to use and why, citing specific M-values and V-values.