Hiscript 3 1st strand cdna synthesis kit

The total RNA was extracted from samples of pepper and Arabidopsis by an RNA extraction kit (Novozymes, Nanjing, China) and measured using a spectrophotometer (Thermo Scientific, Wilmington, DE, USA), then reverse-transcribed to cDNA using HiScript III 1st strand cDNA synthesis kit (Novozymes, Nanjing, China). Primer sequences were designed using Premier 5.0 software (Table S1). As housekeeping genes, CcActin (accession: AY486137.1) from C. chinense and AtActin (accession: NM_001338359.1) from Arabidopsis were utilized. With the aid of the 2ChamQ Universal SYBR qPCR master mix (Novozymes, Nanjing, China), RT-qPCR reactions were carried out and the 2^−ΔΔCT method was ultimately used to process the data.

Total RNA from MDBK or HeLa cells was isolated using a Total RNA Extraction Kit (Tengen, China) according to the manufacturer’s instructions. cDNA was synthesized using a Novozymes HiScript III 1st Strand cDNA Synthesis Kit (Novozymes, Beijing, China). quantitative real-time PCR (qRT-PCR) was performed on a Roche LightCycler 480 II with 2× SYBR Green qPCR Master Mix (Selleck, Beijing, China) according to the manufacturer’s instructions. All qRT-PCR experiments were performed in triplicate. Relative gene levels were determined using the $2^{-\Delta \Delta }Ct$ method with GAPDH as an internal control. The primers used for qRT-PCR are listed in Table 1.