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Sybr greener qpcr supermix

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SYBR GreenER qPCR SuperMix is a ready-to-use solution for quantitative real-time PCR (qPCR) applications. It contains SYBR Green I dye, necessary PCR components, and a DNA polymerase.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (16 mentions) Trizol reagent, by Thermo Fisher Scientific (16 mentions) Rneasy mini kit, by Qiagen (11 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (10 mentions) Abi prism 7900ht, by Thermo Fisher Scientific (9 mentions) 7900ht fast real time pcr system, by Thermo Fisher Scientific (8 mentions) Superscript 3 first strand synthesis system, by Thermo Fisher Scientific (8 mentions) Iscript cdna synthesis kit, by Bio-Rad (8 mentions) Superscript 3 first strand synthesis kit, by Thermo Fisher Scientific (7 mentions) Rneasy micro kit, by Qiagen (7 mentions) Rneasy kit, by Qiagen (4 mentions) Cfx96 real time pcr detection system, by Bio-Rad (4 mentions) Rneasy mini column, by Qiagen (4 mentions) Quantstudio 7 flex real time pcr system, by Thermo Fisher Scientific (4 mentions) Superscript vilo cdna synthesis kit, by Thermo Fisher Scientific (4 mentions) Qiaamp dna mini kit, by Qiagen (3 mentions) Icycler, by Bio-Rad (3 mentions) Abi 7300 real time pcr system, by Thermo Fisher Scientific (3 mentions) 7300 real time pcr system, by Thermo Fisher Scientific (3 mentions) Random primer, by Thermo Fisher Scientific (3 mentions)

Spelling variants of this product

SYBR GreenER (38 citations) SYBR GreenER SuperMix (14 citations) Express SYBR® GreenER qPCRs supermix Universal kit (3 citations)

119 protocols using sybr greener qpcr supermix

1

Quantifying Gene Expression in Tissue Samples

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Total RNA was extracted from frozen tissue using TRIzol (Invitrogen), purified with RNeasy Mini spin columns (QIAGEN) and reverse transcribed using a High-Capacity cDNA Reverse Transcription kit (Applied Biosystems). The resultant cDNA was analyzed by qRT–PCR. Briefly, 20 ng cDNA and 150 nmol of each primer were mixed with SYBR GreenER™ qPCR SuperMix (Applied Biosystems). Reactions were performed in a 384-well format using an ABI PRISM 7900HT real time PCR system (Applied Biosystems). Relative mRNA levels were calculated using the comparative CT method and normalized to cyclophilin mRNA. The following primers were used in these studies: Cyclophilin forward: 5′-GGA GAT GGC ACA GGA GGA A-3′, reverse: 5′-GCC CGT AGT GCT TCA GCT T-3′; Ucp1 forward 5′-ACTGCCACACCTCCAGTCATT-3′, reverse 5′-CTTTGCCTCACTCAGGATTGG-3′; Dio2 forward 5′-CAGTGTGGTGCACGTCTCCAATC-3′, reverse 5′-TGAACCAAAGTTGACCACCAG-3′; Pgc1α forward 5′-CCCTGCCATTGTTAAGACC-3′, reverse 5′-TGCTGCTGTTCCTGTTTTC-3′; PPARγ forward 5′-TGAAAGAAGCGGTGAACCACTG-3′, reverse 5′-TGGCATCTCTGTGTCAACCATG-3′; Pgc1β forward 5′-CTGACGTGGACGAGCTTTCA-3′, reverse 5′-CGTCCTTCAGAGCGTCAGAG-3′; Nrf2 forward 5′-CCAGCTACTCCCAGGTTGCC-3′, reverse 5′-GGGATATCCAGGGCAAGCGA-3′; Ap2 5′-AAGGTGAAGAGCATCATAACCCT-3′, reverse 5′-TCACGCCTTTCATAACACATTCC-3′.
Chouchani E.T., Kazak L., Jedrychowski M.P., Lu G.Z., Erickson B.K., Szpyt J., Pierce K.A., Laznik-Bogoslavski D., Vetrivelan R., Clish C.B., Robinson A.J., Gygi S.P, & Spiegelman B.M. (2016). Mitochondrial ROS regulate thermogenic energy expenditure and sulfenylation of UCP1. Nature, 532(7597), 112-116.
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2

Quantification of miR-142-5p and mRNA Expression

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Total RNA was abstracted from GC cells with RNAiso Plus kit (Takara, Dalian, China). In order to detect miR‐142‐5p expression, total RNA was reversely transcribed with MMLV reverse transcriptase (Takara, Dalian, China). With the cDNA obtained as the template, RT‐PCR was executed with real‐time PCR kit (Qiagen, Shanghai, China). Then the miRNA expression was quantified by real‐time PCR and its relative level was normalized to U6. To detect HCG18 and WIPF1 mRNA expression, first‐strand cDNA was synthesized by using cDNA Synthesis Kit (Takara, Dalian, China) under the guidance of the protocol. PCR was executed by using SYBR greener qPCR SuperMix (Applied Biosystems, Shanghai, China). Relative expression was calculated using 2‐△△Ct method. Table 1 shows the specific primer sequences.
Liu Y., Lin W., Dong Y., Li X., Lin Z., Jia J., Zou W, & Pan Y. (2020). Long noncoding RNA HCG18 up‐regulates the expression of WIPF1 and YAP/TAZ by inhibiting miR‐141‐3p in gastric cancer. Cancer Medicine, 9(18), 6752-6765.
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3

Quantitative Real-Time PCR Analysis of Gene Expression

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Total RNA was extracted by using TRIzol (GIBCO Life Technology). First-strand cDNA was prepared from total RNA (1.5 μg) using the SuperScript® VILO cDNA Synthesis Kit and Master Mix (Thermo Fisher Technologies), and cDNA (1 μl) was amplified in triplicate using SYBR GreenER qPCR Supermix on an ABI Quantstudio® 3 Real-Time PCR instrument (Applied Biosystems). Primers for mouse Klf15, mouse Col1α1, mouse Vimentin, mouse Fibronectin, mouse c-Myc, and mouse Ctgf were designed using NCBI/Primer-BLAST (Supplemental Table 2). Light cycler analysis software was used to determine crossing points using the second derivative method. Data were normalized to housekeeping genes (ACTB) and presented as fold increase compared with RNA isolated from the control group using the 2−ΔΔCT method.
Gu X., Mallipattu S.K., Guo Y., Revelo M.P., Pace J., Miller T., Gao X., Jain M.K., Bialkowska A.B., Yang V.W., He J.C, & Mei C. (2017). The loss of Krüppel-like factor 15 in Foxd1+ stromal cells exacerbates kidney fibrosis. Kidney international, 92(5), 1178-1193.
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4

Quantitative Real-Time PCR Analysis

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Total RNA was extracted by using TRIzol (Invitrogen). First strand cDNA was prepared from total RNA (2.0 μg) using the Superscript ™ III first strand synthesis kit (Invitrogen), and cDNA (1 μl) was amplified in triplicate using SYBR GreenER qPCR Supermix on an ABI PRISM 7900HT (Applied Biosystems, Foster City, CA). The primer sequences are listed in Table 1. Light Cycler analysis software was used to determine crossing points using the second derivative method. Data were normalized to housekeeping genes (GAPDH) and presented as fold increase compared with RNA isolated from WT animals using the 2-ΔΔCT method.
Zhong F., Chen H., Wei C., Zhang W., Li Z., Jain M.K., Chuang P.Y., Chen H., Wang Y., Mallipattu S.K, & He J.C. (2014). Reduced Krüppel-like Factor 2 expression may aggravate the endothelial injury of diabetic nephropathy. Kidney international, 87(2), 382-395.
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5

Quantitative Real-Time PCR Analysis

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Total RNA was extracted by using TRIzol (Invitrogen). First strand cDNA was prepared from total RNA (2.0 μg) using the Superscript ™ III first strand synthesis kit (Invitrogen), and cDNA (1 μl) was amplified in triplicate using SYBR GreenER qPCR Supermix on an ABI PRISM 7900HT (Applied Biosystems, Foster City, CA). The primer sequences are listed in Table 1. Light Cycler analysis software was used to determine crossing points using the second derivative method. Data were normalized to housekeeping genes (GAPDH) and presented as fold increase compared with RNA isolated from WT animals using the 2-ΔΔCT method.
Zhong F., Chen H., Wei C., Zhang W., Li Z., Jain M.K., Chuang P.Y., Chen H., Wang Y., Mallipattu S.K, & He J.C. (2014). Reduced Krüppel-like Factor 2 expression may aggravate the endothelial injury of diabetic nephropathy. Kidney international, 87(2), 382-395.
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6

Quantitative Real-Time PCR analysis

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Total RNA was extracted by using TRIzol (Invitrogen). First strand cDNA was prepared from total RNA (2.0 μg) using the Superscript TM III first strand synthesis kit (Invitrogen), and cDNA (1 μl) was amplified in triplicate using SYBR GreenER qPCR Supermix on an ABI PRISM 7900HT (Applied Biosystems, Foster City, CA). The primer sequences are listed in Supplementary Table 1. Light Cycler analysis software was used to determine crossing points using the second derivative method. Data were normalized to housekeeping genes (GAPDH) and presented as fold increase compared with RNA isolated from control wildtype animals using the 2-∆∆CT method.
Zhong F., Wang W., Lee K., He J.C, & Chen N. (2016). Role of C/EBP-α in Adriamycin-induced podocyte injury. Scientific Reports, 6, 33520.
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7

Quantitative Real-Time PCR Analysis

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First strand cDNA was prepared from total RNA (1.5 ug) using the SuperScript™ III First-Strand Synthesis Kit (Invitrogen) and cDNA (1 μl) was amplified in triplicate using SYBR GreenER qPCR Supermix on an ABI PRISM 7900 HT (Applied Biosystems, Foster City, CA, USA). Primers were designed using PrimerBlast (http://www.ncbi.nlm.nih.gov/tools/primer-blast/) and purchased through Sigma (Table 1). The sequences of the primers are listed in the Table 2. Light cycler analysis software was used to determine crossing points using the second derivative method. Efficiency was calculated for all the designed primers using relative standard curve method. Data was normalized to housekeeping gene (GAPDH) and presented as fold increase compared to RNA isolated from WT animals using the Pfaffl method [32] (link).
Fan Y., Wei C., Xiao W., Zhang W., Wang N., Chuang P.Y, & He J.C. (2014). Temporal Profile of the Renal Transcriptome of HIV-1 Transgenic Mice during Disease Progression. PLoS ONE, 9(3), e93019.
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8

qPCR Analysis of Gene Expression

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Total RNA was extracted by using TRIzol. First-strand cDNA was prepared from total RNA (2.0 μg) using the Superscript III First-Strand Synthesis Kit (Invitrogen) and cDNA (1 μL) was amplified in triplicate using SYBR GreenER qPCR Supermix on an ABI PRISM 7900HT (Applied Biosystems). Light Cycler analysis software was used to determine crossing points using the second derivative method. Data were normalized to a housekeeping gene (Gapdh) and are presented as fold change compared with the control group using the 2–ΔΔCT method. The following primers were used: Wt1 (forward, 5′-GAGAGCCAGCCTACCATCC-3′ and reverse, 5′-GGGTCCTCGTGTTTGAAGGAA-3′), Il6 (forward, 5′-CCAGCTATGAACTCCTTCTC-3′ and reverse, 5′-GCTTGTTCCTCACATCTCTC-3′), and Ccl2 (forward, 5′-AGGTGACTGGGGCATTGAT-3′ and reverse, 5′-GCCTCCAGCATGAAAGTCTC-3′).
Zhong F., Cai H., Fu J., Sun Z., Li Z., Bauman D., Wang L., Das B., Lee K, & He J.C. (2023). TYRO3 agonist as therapy for glomerular disease. JCI Insight, 8(1), e165207.
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9

Mouse Tissue RNA Extraction and qPCR

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Mouse tissue samples were frozen in liquid nitrogen and pulverized using a Bessman tissue pulverizer. Crushed tissue was resuspended in Trizol (Ambion, 15596-018) and processed according to the manufacturer's instructions. After assessing mRNA concentration and quality using a Nanodrop spectrophotometer (ThermoScientific), cDNA was prepared using the High Capacity cDNA Reverse Transcription kit (Applied Biosystems, Part No. 4368813). qPCR was performed (Invitrogen, SYBR GreenER qPCR Supermix, 11762100) according to the manufacturer's specifications using an Applied Biosystems 7900HT Fast Real-Time PCR System (Iowa Institute of Human Genetics). Relative expression was calculated with the ΔΔct method [22] (link) using CycloA as the reference gene.
Cushing E.M., Chi X., Sylvers K.L., Shetty S.K., Potthoff M.J, & Davies B.S. (2017). Angiopoietin-like 4 directs uptake of dietary fat away from adipose during fasting. Molecular Metabolism, 6(8), 809-818.
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10

Quantifying Gene Expression in Tissue Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was extracted from frozen tissue using TRIzol (Invitrogen), purified with RNeasy Mini spin columns (QIAGEN) and reverse transcribed using a High-Capacity cDNA Reverse Transcription kit (Applied Biosystems). The resultant cDNA was analyzed by qRT–PCR. Briefly, 20 ng cDNA and 150 nmol of each primer were mixed with SYBR GreenER™ qPCR SuperMix (Applied Biosystems). Reactions were performed in a 384-well format using an ABI PRISM 7900HT real time PCR system (Applied Biosystems). Relative mRNA levels were calculated using the comparative CT method and normalized to cyclophilin mRNA. The following primers were used in these studies: Cyclophilin forward: 5′-GGA GAT GGC ACA GGA GGA A-3′, reverse: 5′-GCC CGT AGT GCT TCA GCT T-3′; Ucp1 forward 5′-ACTGCCACACCTCCAGTCATT-3′, reverse 5′-CTTTGCCTCACTCAGGATTGG-3′; Dio2 forward 5′-CAGTGTGGTGCACGTCTCCAATC-3′, reverse 5′-TGAACCAAAGTTGACCACCAG-3′; Pgc1α forward 5′-CCCTGCCATTGTTAAGACC-3′, reverse 5′-TGCTGCTGTTCCTGTTTTC-3′; PPARγ forward 5′-TGAAAGAAGCGGTGAACCACTG-3′, reverse 5′-TGGCATCTCTGTGTCAACCATG-3′; Pgc1β forward 5′-CTGACGTGGACGAGCTTTCA-3′, reverse 5′-CGTCCTTCAGAGCGTCAGAG-3′; Nrf2 forward 5′-CCAGCTACTCCCAGGTTGCC-3′, reverse 5′-GGGATATCCAGGGCAAGCGA-3′; Ap2 5′-AAGGTGAAGAGCATCATAACCCT-3′, reverse 5′-TCACGCCTTTCATAACACATTCC-3′.
Chouchani E.T., Kazak L., Jedrychowski M.P., Lu G.Z., Erickson B.K., Szpyt J., Pierce K.A., Laznik-Bogoslavski D., Vetrivelan R., Clish C.B., Robinson A.J., Gygi S.P, & Spiegelman B.M. (2016). Mitochondrial ROS regulate thermogenic energy expenditure and sulfenylation of UCP1. Nature, 532(7597), 112-116.
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