Fast sybr green master mix

Manufactured by Promega

Fast SYBR Green Master mix is a ready-to-use solution for real-time PCR amplification and detection. It contains SYBR Green I dye, DNA polymerase, and necessary reagents for efficient and rapid real-time PCR.

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Lab products found in correlation

Tri reagent, by Merck Group (4 mentions) Gotaq qpcr master mix, by Promega (3 mentions) Ariamx, by Agilent Technologies (3 mentions) Gapdh, by Qiagen (2 mentions) Mb21d1, by Qiagen (2 mentions) Improm 2 reverse transcription system, by Promega (1 mentions) Trizol reagent, by Merck Group (1 mentions) Goscript reverse transcription system, by Promega (1 mentions) Ifnα4, by Qiagen (1 mentions) Primer blast, by Thermo Fisher Scientific (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) M mlv reverse transcriptase, by Promega (1 mentions)

Spelling variants of this product

SYBR Green (122 citations) SYBR Green Mix (24 citations) SYBR master mix (5 citations)

6 protocols using fast sybr green master mix

Lung RNA Extraction and qPCR Analysis

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RNA was purified from lung homogenates using Tri-Reagent (Sigma-Aldrich, Saint-Louis, MO) extraction protocol. RNA reverse transcription into cDNA was carried out with GoTaq qPCR-Master Mix (Promega, Madison, WI). RT-qPCR was performed with Fast SYBR Green Master mix (Promega) on an ARIA MX (Agilent Technologies, Santa Clara, CA). Primers for Fn1 (#QT00135758), Col3a (#QT01055516) and Timp1 (#QT00996282) were purchased from Qiagen (Qiagen, Hilden, Germany). RNA expression was normalized to Gapdh (#QT01658692, Qiagen, Hilden, Germany) expression and analyzed using the ΔΔCt method.

Carignon S., De Moura Rodrigues D., Gosset D., Culerier E., Huot-Marchand S., Savigny F., Kaya E., Quesniaux V., Gombault A., Couillin I., Ryffel B., Le Bert M, & Riteau N. (2023). Lung inflammation and interstitial fibrosis by targeted alveolar epithelial type I cell death. Frontiers in Immunology, 14, 1261483.

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Quantitative Gene Expression Analysis

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Tumor samples were immediately snap‐frozen in liquid nitrogen after harvesting. Total RNA was extracted using TRIzol reagent (#T9424, Sigma‐Aldrich, St. Louis, MO, USA). Complementary DNA was synthesized from 2 μg total RNA with the ImProm‐II Reverse Transcription System (#A3800, Promega, Madison, WI, USA), according to the manufacturer's directions. Real‐time PCR analysis was carried out with diluted cDNA, and Fast SYBR Green Master Mix (#4385616, Promega) was used. The sequences of the primers were as follows.
CD8A: forward, 5′‐TCCTCCTATACCTCTCCCAAAAC‐3′ – reverse, 5′‐GGAAGACCGGCACGAAGTG‐3′; IFNG: forward, 5′‐TCGGTAACTGACTTGAATGTCCA ‐3′ – reverse, 5′‐TCGCTTCCCTGTTTTAGCTGC‐3′; IDO1: forward, 5′‐TCTCATTTCGTGATGGAGACTGC‐3′ reverse, 5′‐GTGTCCCGTTCTTGCATTTGC‐3′; GAPDH forward: 5′‐GGAGCGAGATCCCTCCAAAAT‐3′ reverse, 5′‐GGCTGTTGTCATACTTCTCATGG‐3′.
Expression relative to the reference gene GAPDH was calculated as 2−ΔCt , where ΔC_t is Ctgene‐Ctctrl. If undetectable, Ct was given a value of 40.

Li S., Han X., Lyu N., Xie Q., Deng H., Mu L., Pan T., Huang X., Wang X., Shi Y, & Zhao M. (2018). Mechanism and prognostic value of indoleamine 2,3‐dioxygenase 1 expressed in hepatocellular carcinoma. Cancer Science, 109(12), 3726-3736.

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RNA Extraction and RT-qPCR Analysis

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RNA was purified from lung homogenates by using Tri-Reagent (Sigma-Aldrich) extraction protocol. Reverse transcription of RNA into cDNA was carried out with GoScript™ Reverse Transcription System (Promega). RT-qPCR was performed with Fast SYBR Green Master mix (Promega) on anARIAMX(Agilent Technologies). Primers for Tnfsf13 (#QT00254023) were purchased from Qiagen (Qiagen, Hilden, Germany). RNA expression was normalized to Gapdh (#QT00166768) expression and analyzed using the ^ΔΔCt method.

Nascimento M., Huot-Marchand S., Gombault A., Panek C., Bourinet M., Fanny M., Savigny F., Schneider P., Le Bert M., Ryffel B., Riteau N., Quesniaux V.F, & Couillin I. (2020). B-Cell Activating Factor Secreted by Neutrophils Is a Critical Player in Lung Inflammation to Cigarette Smoke Exposure. Frontiers in Immunology, 11, 1622.

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qRT-PCR of STING, cGAS, and Fibronectin

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RNA was purified from lung homogenates using Tri-Reagent (Sigma-Aldrich, Saint-Louis, MO) extraction protocol. RNA reverse transcription into cDNA was carried out with GoTaq qPCR-Master Mix (Promega, Madison, WI). RT-qPCR was performed with Fast SYBR Green Master mix (Promega) on an ARIA MX (Agilent Technologies, Santa Clara, CA). Primers Tmem173 (#QT00261590) encoding for STING, Mb21d1 (#QT00131929) encoding for cGAS, and Fn1 (#QT00135758) encoding for fibronectin were purchased from Qiagen (Qiagen, Hilden, Germany). RNA expression was normalized to Gapdh (#QT00166768, Qiagen, Hilden, Germany) expression and analyzed using the ΔΔCt method.

Savigny F., Schricke C., Lacerda-Queiroz N., Meda M., Nascimento M., Huot-Marchand S., Da Gama Monteiro F., Ryffel B., Gombault A., Le Bert M., Couillin I, & Riteau N. (2021). Protective Role of the Nucleic Acid Sensor STING in Pulmonary Fibrosis. Frontiers in Immunology, 11, 588799.

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Lung RNA Expression Analysis

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RNA was purified from lung homogenates by using Tri-Reagent (Sigma-Aldrich, Saint-Louis, MO) extraction protocol. Reverse transcription of RNA into cDNA was carried out with GoTaq qPCR-Master Mix (Promega, Madison, WI). RT-qPCR was performed with Fast SYBR Green Master mix (Promega) on an ARIA MX (Agilent Technologies, Santa Clara, CA). Primers for Tmem173 (#QT00261590), Mb21d1 (#QT00131929) and Ifnα4 (#QT01774353) were purchased from Qiagen (Qiagen, Hilden, Germany). RNA expression was normalized to Gapdh (#QT00166768) expression and analysed using the ^ΔΔCt method.

Nascimento M., Gombault A., Lacerda-Queiroz N., Panek C., Savigny F., Sbeity M., Bourinet M., Le Bert M., Riteau N., Ryffel B., Quesniaux V.F, & Couillin I. (2019). Self-DNA release and STING-dependent sensing drives inflammation to cigarette smoke in mice. Scientific Reports, 9, 14848.

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Total RNA Extraction and Real-Time qPCR Analysis

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Total RNA was extracted with Trizol (Invitrogen). cDNA was synthesized with M-MLV reverse transcriptase (Promega) and quantified by realtime qPCR using Biosystems StepOne™ Real-Time PCR system and Fast SYBR Green Master Mix (Promega). PCR primers were designed with NCBI online software Primer-BLAST and synthesized by Invitrogen. The sequences of primers used in this study are listed in Table 1.

He H., Du F., He Y., Wei Z., Meng C., Xu Y., Zhou H., Wang N., Luo X.G., Ma W, & Zhang T.C. (2018). The Wnt-β-catenin signaling regulated MRTF-A transcription to activate migration-related genes in human breast cancer cells. Oncotarget, 9(20), 15239-15251.

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