Total RNA was extracted from liver tissue by Trizol reagent (CatNo. R1100, Solarbio) and was reversely transcribed to cDNA according to the manufacturer’s protocol (HiScript II Q RT SuperMix, CatNo. R223-01, Solarbio). Primers were designed and purchased from Solarbio and quantitative PCR was further performed following the manufacturer’s protocol (Novozan AceQ qPCR SYBR GreenMaster Mix, CatNo. Q111-02, Solarbio) using LightCycler96 real-time fluorescence quantitative PCR instrument (CatNo. 05815916001, Roche)(Supplementary Table 1
Hiscript 2 q rt supermix
Manufactured by Solarbio
HiScript II Q RT SuperMix is a ready-to-use reagent for reverse transcription and real-time quantitative PCR (RT-qPCR) experiments. It contains a high-performance reverse transcriptase and a hot-start Taq DNA polymerase, optimized for sensitive and specific gene expression analysis.
Automatically generated - may contain errors
2 protocols using hiscript 2 q rt supermix
1
Quantitative Analysis of Liver RNA Expression
2
Quantitative Analysis of mRNA and miRNA Expression
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Total RNA was extracted from the liver tissues and AML12 cells using the Triquick reagent (cat. no. R1100; Beijing Solarbio Science & Technology, Inc.) according to the manufacturer's instructions, and reverse transcribed into cDNA using the HiScript II Q RT SuperMix (cat. no. R223-01; Vazyme Biotech Co., Ltd.) or miRNA First Strand cDNA Synthesis (cat. no. B532453; Sangon Biotech Co., Ltd.). qPCR analysis was performed using the ChamQ Universal SYBR qPCR Master Mix (cat. no. Q711-02; Vazyme Biotech Co., Ltd.) or miRNA Universal SYBR qPCR Master Mix (cat. no. MQ101-02; Vazyme Biotech Co., Ltd.) according to the manufacturer's instructions. The thermocycling conditions were 95°C for 30 sec, 95°C for 5 sec, and 60°C for 35 sec (40 cycles). The expression of mRNA and miRNA was normalized to the expression of GAPDH and U6, respectively. Fold change was calculated by the 2−∆∆Cq method (32 (link)). The sequences of the primers used in this experiment are provided in Table I .
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