Real-Time PCR Analysis of GmCYP78A Genes

Total RNAs were isolated from plant tissues with Trizol (TIANGEN Biotech, Beijing, China) following the manufacturer’s instructions. RNA quality was determined by a Nanophotomoter (Implen, München, Germany). The removal of genomic DNA residues, reverse transcription and cDNA synthesis were separately performed with 2 μg of total RNA and a FastQuant cDNA RT Kit (TIANGEN Biotech). Real-time PCR analysis of GmCYP78As was performed with the FastStart Essential DNA Green Master (Roche, Shanghai, China) on a Stratagene Mx3005P (Agilent Technologies, Santa Clara, CA, USA, Supplementary Table S1). The relative expression levels were calculated from three replicates using the 2^−ΔΔCt method after normalization to the Actin11 control in soybean [31 (link)].