Superscript 2 reverse transcriptase first strand synthesis kit

All mungbean RNA was extracted using Qiagen RNeasy mini kit following the instructions (Qiagen, United States)¹¹. cDNA synthesis was conducted as previously described (Li et al., 2017 (link)). SuperScript II reverse transcriptase first-strand synthesis kit (Invitrogen) was used for the synthesis of the first strand cDNAs with 2 μg total RNA according to the instructions. The LightCycler 480 SYBR Green I Master Kit (Roche Diagnostics) was used for quantitative real-time PCR (qRT-PCR) using a LightCycler480 machine (Roche Diagnostics), according to the manufacturer’s instructions. The amplification program for qRT-PCR was performed as previously described (Li et al., 2017 (link)). For gene expression analysis, three biological replicates were used for each sample and gene expression was normalized to an Actin-expressing gene in mungbean (Vradi03g00210). All primers used for qRT-PCR analyses are listed in Supplementary Table 1.