Figure 8

Microarray data validation by qRTPCR. Specific primers were designed to amplify exclusively the oligo sequences found by microarray data and confirmed by RNA gel blot analysis. The two clones chosen were the Auxin down regulated ADR12-2 and the Chlorophyll a/b binding protein. All reactions were conducted in triplicate starting with purified total RNA (5 μg) from each developmental stage (S1–S7) from one biological replicate that was converted into first strand cDNA by reverse transcription. PCR product quantification was based on a Ct value. The relative expression of each stage (S2–S7) compared to the reference imbibed seed tissue (S1) was calculated by averaging the 3 Ct values for each measurement and determine the relative ratio using the 2^-ΔΔCt method [62].