Miscript syber green master mix

Manufactured by Qiagen

Sourced in Germany

The MiScript Syber green master mix is a reagent used for quantitative real-time PCR (qRT-PCR) analysis. It contains SYBR Green I dye, which binds to double-stranded DNA and emits fluorescence, enabling the detection and quantification of target gene expression.

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Lab products found in correlation

Miscript rt kit, by Qiagen (2 mentions) 5 plex rotor gene pcr analyzer, by Qiagen (2 mentions) Mireasy mini kit, by Qiagen (2 mentions) Miscript primer assay, by Qiagen (1 mentions) Quantitect sybr green pcr kit, by Qiagen (1 mentions)

2 protocols using miscript syber green master mix

Quantification of miRNA and mRNA Expression

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As per the manufacturer’s protocol, total mRNA and miRNAs were extracted with a miReasy Mini Kit (Qiagen, Hilden, Germany). The complementary DNA (cDNA) was prepared via a reverse transcription reaction from miScript RT Kit (Qiagen, Hilden, Germany). The miRNA and mRNA genes were amplified via miScript Syber green master mix and QuantiTect SYBR Green PCR Kit, respectively (Qiagen, Hilden, Germany). The 5 plex Rotor-Gene PCR Analyzer (Qiagen, Hilden, Germany).
The relative expression level (fold change) for the miR-let7a and HO-1 gene was normalized to SNORA11 and β-actin internal controls, respectively, and relative to the calibrator (negative control sample), they were calculated using the equation 2^−∆∆Ct test [48 (link),49 (link)]. The relative gene expression of miR-let7a and HO-1 was determined using the miScript primer assay (miR-let-7a), catalogue number: MS00031220, and QuantiTect primer assay (HO-1), catalogue number: 249900 (Qiagen, Germany). Regarding the housekeeping genes, SNORA11, miScript primer assay with catalogue number 218300 and β-actin gene, ACTB_ 1_SG QuantiTect primer assay with catalogue number 249900, were used (Qiagen, Hilden, Germany).

Attallah N.G., Kabbash A., Negm W.A., Elekhnawy E., Binsuwaidan R., Al-Fakhrany O.M., Shaldam M.A., Moglad E., Tarek M., Samir N, & Fawzy H.M. (2023). Protective Potential of Saussurea costus (Falc.) Lipsch. Roots against Cyclophosphamide-Induced Pulmonary Injury in Rats and Its In Vitro Antiviral Effect. Pharmaceuticals, 16(2), 318.

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Quantification of miRNA-155 and miRNA-21 expression

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Total mRNA was extracted using miReasy Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's protocol. cDNA was synthesized by reverse transcription reaction using mi Script RT Kit (Qiagen, Hilden, Germany). The quantification of miRNA-155 and miRNA-21 genes expression levels was amplified from miRNA extract using a miScript (Hs_miR-21 and miR-155a) Primer Assays. The miRNAs genes were amplified by miScript Syber green Master mix (Qiagen, Hilden, Germany). The RUN6 Primer Assay was used as a housekeeper gene. All samples were analyzed using the 5 plex Rotor-Gene PCR Analyzer (Qiagen, Germany).

mohamed L.A., Bolok A.H.M.E., Elgayar S.F., & Fahmy A.A. (2022). miRNA-155 as a Novel Target for Isoliquiritigenin to Induce Autophagy in Oral Squamous Cell Carcinoma. Open Access Macedonian Journal of Medical Sciences, 10(A), 481-488.

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