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Hiscript 3 supermix for qpcr

Manufactured by Vazyme
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Sourced in China

HiScript® III SuperMix for qPCR is a ready-to-use reaction mix designed for quantitative real-time PCR (qPCR) applications. It contains all the necessary components, including a high-performance reverse transcriptase and a hot-start DNA polymerase, to facilitate efficient cDNA synthesis and amplification in a single tube.

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Lab products found in correlation

Cfx96 thermal cycler, by Bio-Rad (2 mentions) Fastpure cell tissue total rna isolation kit, by Vazyme (2 mentions) Itaq master sybr green super mix, by Bio-Rad (2 mentions) Sv total rna isolation system kit, by Promega (1 mentions) Iq5 pcr system, by Bio-Rad (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Chamq universal sybr qpcr master mix, by Vazyme (1 mentions)

Spelling variants of this product

HiScript III RT SuperMix for qPCR (259 citations) HiScript III RT SuperMix for qPCR kit (81 citations) HiScript III All-in-one RT SuperMix Perfect for qPCR Kit (27 citations) HiScript III RT SuperMix for qPCR with gDNA wiper (12 citations) HiScript III SuperMix (4 citations) HiScript III (4 citations) HiScript® III SuperMix for qPCR (+gDNA wiper) (4 citations) HiScript III RT SuperMix for RT-qPCR (4 citations) HiScript® III All-in-one RT SuperMix Perfect for RT-qPCR (3 citations) HiScript III RT SuperMix 100 for qPCR (+gDNAwiper) (2 citations)

4 protocols using hiscript 3 supermix for qpcr

1

Quantitative Gene Expression Analysis in Lung Tissue

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The total RNA of lung tissues was extracted using a FastPure® Cell/Tissue Total RNA Isolation Kit (Vazyme Biotech Co., Ltd., Nanjing, China) following the manufacturer’s instructions. First-strand cDNA was synthesized using HiScript® III SuperMix for qPCR (Vazyme Biotech Co., Ltd., Nanjing, China), and stored at −20 °C. A real-time PCR reaction system was prepared using the iTaq Master SYBR Green Super Mix (Bio-Rad Laboratories Inc., Hercules, CA, USA) and carried out with a CFX96 Thermal Cycler (Bio-Rad Laboratories Inc.). The expressions of genes were calculated using the 2−ΔΔCT method, and normalized to that of GAPDH. Table 1 shows the primer sequences.
Wang Q., Fang Z., Xiao Y., Wang H., Zhang P., Lu W., Zhang H, & Zhou X. (2023). Lactiplantibacillus pentoses CCFM1227 Produces Desaminotyrosine to Protect against Influenza Virus H1N1 Infection through the Type I Interferon in Mice. Nutrients, 15(16), 3659.
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2

RNA Extraction and qRT-PCR Analysis

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Total RNA was extracted from lung tissues using the FastPure® Cell/Tissue Total RNA Isolation Kit (Vazyme Biotech Co., Ltd., Nanjing, China). First-strand cDNA was synthesized from total RNA using HiScript® III SuperMix for qPCR (Vazyme Biotech Co., Ltd., Nanjing, China). Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis was performed using iTaq Master SYBR Green Super Mix (Bio-Rad Laboratories Inc., Hercules, CA, United States) and a CFX96 Thermal Cycler (Bio-Rad Laboratories Inc.). The relative expression of genes was normalized to that of GAPDH and calculated using the 2−ΔΔCT method (Livak and Schmittgen, 2001 (link)). Table 1 shows the primer sequences.
Wang Q., Fang Z., Li L., Wang H., Zhu J., Zhang P., Lee Y.K., Zhao J., Zhang H., Lu W, & Chen W. (2022). Lactobacillus mucosae exerted different antiviral effects on respiratory syncytial virus infection in mice. Frontiers in Microbiology, 13, 1001313.
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3

Quantitative RT-PCR Analysis of Bacterial Gene Expression

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RNA samples were isolated from fresh bacterial cultures (OD600 = 1.0) using the SV total RNA isolation system kit (Promega). The integrity of RNA was visualized by agarose gel electrophoresis and the concentration of RNA was measured using a NanoDrop ND-100 spectrophotometer.
Reverse transcription PCR was performed using a HiScript III SuperMix for qPCR (Vazyme) according to the manufacturer’s instructions. Specific RT-qPCR primers listed in Table S1 were used to amplify central coding fragments of approximately 200 bp in length from different genes. The quality of primers for amplification capability was determined by the melting curve analysis. SYBR Green qPCR Master Mixes (Vazyme) was used according to the manufacturer’s instructions. As a control, RT-qPCR was similarly performed to analyze arcA gene expression. The absolute value of −ΔΔCt = −(ΔCt1 − ΔCt2) was calculated as described in the formula 2−ΔΔCt (Livak and Schmittgen, 2001 (link)). The RT-qPCR experiment was repeated at least twice and the cDNA samples were prepared from triplicate cultures each time.
Lv M., Ye S., Hu M., Xue Y., Liang Z., Zhou X., Zhang L, & Zhou J. (2022). Two-component system ArcBA modulates cell motility and biofilm formation in Dickeya oryzae. Frontiers in Plant Science, 13, 1033192.
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4

Circadian Rhythm and Lipid Metabolism Regulation

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According to the instructions, total RNA was extracted with TRIzol reagent (Invitrogen, USA), RNA was reverse transcribed into cDNA by HiScript ®III SuperMix for qPCR (Vazyme, China), and ChamQ Universal SYBR qPCR Master Mix (Vazyme, China) and the IQ5 PCR System were used (Bio-Rad, USA) to analyze the relative expression of mRNA. The primer sequences used were as follows: Per2 forward, 5’-TTGGACAGCGTCATCAGGTA-3’ and reverse, 5’-TCCGCTTATCACTGGACCTT-3’; SREBP1 forward, 5’-GACAGCCCAGTCTTTGAGGA-3’ and reverse, 5’-CAGGACAGGCAGAGGAAGAC-3’; FASN forward, 5’-CACAGGGACAACCTGGAGTT-3’ and reverse, 5’-ACTCCACAGGTGGGAACAAG-3’; SCD forward, 5’-CGACGTGGCTTTTTCTTCTC-3’ and reverse, 5’-CCTTCTCTTTGACAGCTGGG-3’; ACACA forward, 5’-AGTGGGTCACCCCATTGTT-3’ and reverse, 5’TTCTAACAGGAGCTGGAGCC-3’; Bmal1 forward, 5’-CTCCTCCAATGTGGGCATCAA-3’, and reverse, 5’-GGTGGCACCTCTTAATGTTTTCA-3’; CLOCK forward, 5’-TGCGAGGAACAATAGACCCAA-3’ and reverse, 5’-ATGGCCTATGTGTGCGTTGTA-3’; Cry1 forward, 5’-CTCCTCCAATGTGGGCATCAA-3’ and reverse, 5’-CCACGAATCACAAACAGACGG-3’; Cry2 forward, 5’-TCCCAAGGCTGTTCAAGGAAT-3’ and reverse, 5’-TGCATCCCGTTCTTTCCCAAA-3’; PER1 forward, 5’-AGTCCGTCTTCTGCCGTATCA-3’ and reverse, 5’-AGCTTCGTAACCCGAATGGAT-3’; PER3 forward, 5’-GCAGAGGAAATTGGCGGACA-3’ and reverse, 5’-GGTTTATTGCGTCTCTCCGAG-3’.
Yao J., Hui J.W., Chen Y.J., Luo D.Y., Yan J.S., Zhang Y.F., Lan Y.X., Yan X.R., Wang Z.H., Fan H, & Xia H.C. (2023). Lycium barbarum glycopeptide targets PER2 to inhibit lipogenesis in glioblastoma by downregulating SREBP1c. Cancer Gene Therapy, 30(8), 1084-1093.
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