Figure 2.

Validation of candidate reference genes in Barnea, Frantoio and Picual as a whole using GeNorm algorithm in qBase Plus. (A) Average expression stability values (M) of the eight reference genes plotted from least stable (left) to most stable (right). The M value was calculated for each gene and the least stable gene with the highest M value was excluded from the next calculation round. (B) Pairwise variation analysis (V_n/V_n+1) between the normalisation factors NFn and NFn + 1 to determine the optimal number of reference genes to be used for normalisation against target genes. GeNorm V calculates the normalisation factor (NF_n) by calculating the geometric mean of the expression levels of the stable most reference genes by step-wise inclusion of a less stable gene.