Table 1.
Candidate reference genes and its primer sequences.
Fig 1.
A. Primer specificity and amplicon size. Agarose gel electrophoresis (2.0%) indicates amplification of a single PCR product of the expected size for 10 genes (Line 1–10: ACTIN, EF, SLEEPER, GAPDH1, GAPDH2, TUB, UBI, DSK2A, CYC, and PLA). B. Melting curve analysis: Melting curves of 10 genes show single peaks.
Fig 2.
The quantification cycle (Cq) values of the candidate reference genes in staminate, pistillate and total samples.
Lines across the Box-plot graph of Cq values represent the median values. Lower and upper boxes show the 25th percentile to the 75th percentile. The whisker caps represent the maximum and minimum values.
Fig 3.
Gene expression stability values (M) and ranking of 10 reference genes as assayed by GeNorm in staminate, pistillate and total samples.
The least stable genes are on the left and the most stable genes on the right.
Fig 4.
Pairwise variation (V) of the candidate reference genes calculated by geNorm in staminate, pistillate and total samples.
Vn/Vn+1 value were used for decision of the optimal number of reference genes.
Table 2.
Expression stability analysis of reference genes assayed by NormFinder software.
Table 3.
Expression stability analysis of reference genes assayed by BestKeeper software.
Table 4.
Expression stability analysis of reference genes assayed by ΔCt method.
Table 5.
Most stable and least stable reference genes based on RefFinder analysis.
Fig 5.
Relative expression of AGAMOUS gene using selected reference genes including most stable and least stable reference genes for normalization in staminate and pistillate flowers.
The error bars represent standard deviation.
Fig 6.
The copy number was calculated by standard curve at different developmental stages for staminate and pistillate flower samples of Jatropha curcas.