Transtaq t dna polymerase

An EasyPure Plant RNA Kit (TransGen) was used to isolate total RNA from each frozen sample and the first-strand cDNA was synthesized from total RNA (1 μg) by using EasyScript One-Step gDNA Removal and cDNA Synthesis SuperMix (TransGen) according to the manufacturer’s instructions. The sequence was amplified using gene-specific primers (Table S7) with TransTaq-T DNA Polymerase (TransGen) and the actin gene was used as an internal control. The real-time PCR cycling parameters were 94 °C for 30 s followed by 45 cycles at 94 °C for 5 s and 55 °C for 30 s with a melting curve analysis from 60 °C to 90 °C at a rate of 0.5 °C/5 s. All reactions were performed in triplicate to ensure the reproducibility of the results.

SteadyPure Universal RNA Extraction Kit (Accurate Biology) was used to isolate total RNA, and the first-strand cDNA was synthesized by using the Evo M-MLV kit with gDNA clean for qPCR II (Accurate Biology). The specific primers of HvZF-HD and HvActin genes were showed in Table S1 [26 (link)]. Real-time PCR experiments were conducted using TransTaq-T DNA Polymerase (TransGen, Beijing, China) under the following cycling program: initial denaturation at 95 °C for 3 min, followed by 40 cycles of denaturation at 95 °C for 5 s and annealing/extension at 60 °C for 30 s. Real-time PCR experiments were performed in three independent biological replicates and three technical replications to determine the average Ct values. The relative expression levels of HvZF-HD genes were calculated by 2^−∆∆CT method [27 (link)].

RNApure Plant Kit (CWBIO) was used to isolate total RNA from each frozen sample, and first-strand cDNA was synthesized from total RNA (1 μg) by using PrimeScript™ II 1st Strand cDNA Synthesis Kit (TaKaRa) according to the manufacturer’s instructions. The sequence was amplified using gene-specific primers (Table S2) with TransTaq-T DNA Polymerase (TransGen), and the Actin gene was used as an internal control. The real-time PCR cycling parameters were 94 °C for 30 s, followed by 45 cycles at 94 °C for 5 s and 55 °C for 30 s, with a melting curve analysis from 60 °C to 90 °C at a rate of 0.5 °C/5 s. All reactions were performed in triplicate to ensure the reproducibility of the results.