Sybr green pro taq hs premix 2 kit

The head kidney total mRNA was obtained using an RNA extraction kit (TansGen Biotech, Co., Ltd. (Beijing, China)). The complementary DNA (cDNA) was transcribed using an Evo M-MLV RT kit (Accurate Biotechnology (Accurate Biotechnology Co., Ltd. (Changsha, China)).
The gene expressions of toll-like receptor 2 (TLR2), myeloid differentiation primary response gene 88 (myd88), interleukin 1β (IL-1β), interleukin 10 (IL10), interleukin 8 (IL8) were performed on Roche Light Cycler 480Ⅱ (Switzerland) using an SYBR^® Green Pro Taq HS Premix II kit (Accurate Biotechnology Co., Ltd. (Changsha, China)). The primer sequences of genes are shown in Table 2. The results of real-time qPCR were analyzed by previous authors [33 (link)].

Total RNA samples were extracted from cultured BMMSCs using the Total RNA Extraction Kit (#15596018; GIBCO, USA), as per the manufacturer's instructions. Using the NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, USA) detected the RNA concentrations. The cDNAs were subsequently synthesized from 1 μg total RNA using the Evo M‐MLVRT Kit (#AG11706; Accurate Biotechnology, Hunan, China). Then, a qRT‐PCR assay was performed using the SYBR Green Pro Taq HS Premix II Kit (#AG11702; Accurate Biotechnology, Hunan, China), as per the manufacturer's instructions. The mRNA expression levels were calculated using the standard 2^−ΔΔCt method based on at least three biological replicates. The primer sequences of genes involved in this manuscript were shown in Table 1.

All mRNA expression in total RNA was extracted from the liver using RNA Extraction Kit (TansGen Biotech, Co., Ltd., Beijing, China), and the RNA concentration was determined by nuclear acid protein analyzer (Guangzhou Yitao Scientific Instrument Corporation, Guangzhou, China). Complementary DNA (cDNA) was synthesized by Evo M-MLV RT kit (Accurate Biotechnology (Hunan) Co., Ltd., Hunan, China). All gene expressions of interleukin-6 (IL6), catalase (CAT) and SOD were performed on Roche Light Cycler 480II (Switzerland) using an SYBR^® Green Pro Taq HS Premix II kit (Accurate Biotechnology (Hunan) Co., Ltd., Hunan, China). Primer sequences were designed using Primer 5 following the cDNA sequences found in NCBI (Table 2). The results of real-time qPCR were analyzed by the 2^−ΔΔCT method [18 (link)].