The total RNA of the materials has been extracted using a TransZol Up reagent kit (Transgen, Beijing, China). The RNA quality was analyzed with a Nanodrop 2000 spectrophotometer (Thermo Scientific, Wilmington, DE, USA). The first-strand cDNA was synthesized with a PrimeScript™ IV 1st Strand cDNA Synthesis Mix Kit (TaKaRa, Beijing, China) for qRT-PCR. The DNAMAN software was used to design the primers of the qRT-PCR (Table 1 ), and the qRT-PCR (Roche Lightcyler 480, Roche, Basel, Switzerland) used was SYBR Green chemistry (2 × SYBR Green PCR Master Mix, Xiamen Biocontrast SciTech Co, Ltd., Xiamen, China). DlFeSOD (EU330204) was used as a reference gene for internal control [37 (link)]. The operating parameters of the qRT-PCR are as follows: 95 ℃ for 30 s, followed by 40 cycles of 95 ℃ for 10 s and 58 ℃ for 30 s. The relative expression of the genes involved in PA biosynthesis and metabolism were calculated via the 2− ΔΔCt method [37 (link)].
Transzol up reagent kit
Manufactured by Transgene
Sourced in China
TransZol Up is a reagent kit designed for the extraction and purification of RNA from a variety of biological samples. The kit utilizes a guanidinium thiocyanate-phenol-based solution to facilitate the isolation of high-quality RNA.
Automatically generated - may contain errors
Lab products found in correlation
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Primescript 4 1st strand cdna synthesis mix kit, by Takara Bio (1 mentions)
Lightcyler 480, by Roche (1 mentions)
Transscript one step gdna removal and cdna synthesis supermix kit, by Transgene (1 mentions)
Stepone real time pcr system, by Thermo Fisher Scientific (1 mentions)
Superreal premix plus sybr green kit, by Tiangen Biotech (1 mentions)
2 protocols using transzol up reagent kit
1
Quantitative Expression Analysis of PA Biosynthesis
2
Quantifying Cold-Induced Rice Gene Expression
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Total RNA was extracted from Taipei309 seedling leaves grown under normal (control) conditions or under cold treatment (4 ~ 6 °C for 1 ~ 4 days) using a TransZol Up Reagent Kit according to the manufacturer's protocol (TransGen Biotech, China). The rst stand cDNA synthesis was performed using TransScript® One-Step gDNA Removal and cDNA Synthesis SuperMix Kit (TransGen Biotech, China). SuperReal PreMix Plus (SYBR Green) Kit (TIANGEN, China) was used for qRT-PCR analysis and carried out on a StepOne Real-Time PCR System (Applied Biosystems, USA). Real-time PCR was nished with OsAnn5-F and OsAnn5-R gene-speci c primers (Table S1) as described previously (Shen et al. 2014) . The relative expression level was evaluated using means from three biological samples with three technical replicates, and the ampli cation of the ubiquitin gene (Os03g0234200) was used as an internal control for normalizing all data.
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