Hiscript 2 q rt supermix for qpcr gdna wiper synthesis kit

Total RNA was extracted using the Trizol reagent. The cDNA was synthesized following the instructions of the HiScript® II Q RT SuperMix for qPCR (+gDNA wiper) synthesis kit (Vazyme, Nanjing, China). qPCR was performed on a Step One Plus™ Real-Time PCR system (Applied Biosystems, USA) using ChamQ Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China). Three independents biological replicates were analyzed. Samples used for RNA extraction were collected from plant roots treated as mentioned above.

To validate RNA-seq data, ion-transport-related genes Cluster−3509.57758, Cluster−3509.35440, Cluster−3509.102694, Cluster−3509.89148, and Cluster−3509.97678, and auxin-related genes Cluster−3509.33603 and Cluster−3509.66285 were used as candidate genes. The total RNA was extracted using a FastPure Universal Plant Total RNA Isolation Kit (Vazyme, Nanjing, China). The cDNA was synthesized using a HiScript^® II Q RT SuperMix for qPCR (+gDNA wiper) synthesis kit (Vazyme, China). RT-PCR was performed on BIO-RAD CFX-Opus 96 (Bio-Rad, Hercules, CA, USA) using ChamQ Universal SYBR qPCR Master Mix (Vazyme, China) [3 (link)]. Three biological replicates were analyzed and actin was used for normalization.