Sybr green rt pcr reagent kit luna universal qpcr master mix

Total RNA was extracted from leaves using a Nucleospin RNA Plant Kit (Macherey-Nagel, Duren, Germany). Reverse transcription was performed using a Stratagene Reverse Transcription Kit (Stratagene, La Jolla, CA, USA). Real-time PCR (qRT-PCR) was performed using a Mic qPCR Cycler System (Bio Molecular Systems, Queensland, Australia) using a SYBR Green RT-PCR Reagent Kit (Luna Universal qPCR Master Mix; NEB, Hitchin, UK) according to the manufacturer’s protocol. We used Cyclophilin (Cyclo) or Nt16s rRNA as a normalization control for qRT-PCR and reverse-transcription (RT)-PCR. The RT-PCR conditions were as described previously in [30 (link)]. The primer sequences for RNA expression analysis are listed in Table S1.

Total RNA was extracted from Arabidopsis plants using a Nucleospin RNA Plant Kit (Macherey-Nagel, Duren, Germany). Reverse transcription was performed using a Stratagene Reverse Transcription Kit (Stratagene, La Jolla, CA, USA) with 500 ng of oligo (dT)18 or random octamer primer for RBCL and ClpP1 (CancerROP, Seoul, Korea). The PCR reaction was conducted as following conditions initial denaturation 95 °C (3 min), denaturation 95 °C (30 s), annealing 56 °C (30 s), and extension 72 °C (1 min) with 30 µL of master mix. qRT-PCR was performed on a Mic qPCR Cycler System (Bio Molecular Systems, Queensland, Australia) using SYBR Green RT-PCR Reagent Kit (Luna Universal qPCR Master Mix; NEB, Hitchin, UK) in accordance with the manufacturer’s protocol. The primer sequences for RNA analysis are shown in Table S1. EF-1α (EF1ALPHA; At5g60390) was used for signal normalization. The data were analyzed by analysis of variance using IBM SPSS Statistics 25 software (IBM Corp. Armonk, NY, USA). Means with different letters or asterisks indicate significantly different values at p < 0.05 according to a post hoc Tukey’s honestly significant difference (HSD) test. All data are presented as mean ± standard deviation.