DNA, RNA extraction and cDNA synthesis

Ten days seedlings were ground in liquid nitrogen. Genomic DNA was extracted using CTAB method (Doyle 1991). Total RNA was extracted using RNxplus RNA extraction buffer (SinaClon BioScience, Tehran, Iran) according to manufacturer’s instructions. The quantity and quality of the extracted nucleic acids were determined using nanodrop and agarose gel electrophoresis. Genomic DNA was removed from total RNAs using DNase I (Thermo Fisher Scientific, Lenexa, USA). First-strand cDNA synthesis was performed on 2 µg of total RNA using RevertAid first strand cDNA synthesis kit (Thermo Fisher Scientific, Lenexa, USA). The successful synthesis of cDNA was confirmed by RT-PCR using specific elongation factor 1 alpha (EF1a) primers (Table 1).

Primers used for gene isolation