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Sybr green fastmix

Manufactured by Quanta Biosciences
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Sourced in Canada, Germany

SYBR Green FastMix is a ready-to-use solution for real-time PCR, containing all the necessary components for amplification and detection, including a proprietary hot-start DNA polymerase, SYBR Green dye, and optimized buffer system.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (9 mentions) Qscript cdna supermix, by Quanta Biosciences (5 mentions) Iscript cdna synthesis kit, by Bio-Rad (3 mentions) Rna to cdna kit, by Thermo Fisher Scientific (3 mentions) Cfx96 touch real time pcr detection system, by Bio-Rad (3 mentions) Trizol, by Thermo Fisher Scientific (2 mentions) Ab8580, by Abcam (2 mentions) Ab4729, by Abcam (2 mentions) Protease inhibitor cocktail, by Roche (2 mentions) Cfx96 real time pcr detection system, by Bio-Rad (2 mentions) Rneasy plus mini kit, by Qiagen (2 mentions) Rneasy mini kit, by Qiagen (2 mentions) Rotor gene q, by Qiagen (2 mentions) Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) Qscript cdna synthesis kit, by Quanta Biosciences (2 mentions) Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) Superscript 2, by Thermo Fisher Scientific (1 mentions) Abi prism 7900 sequence detection system, by Thermo Fisher Scientific (1 mentions) Quantitect primer assay, by Qiagen (1 mentions) Superscript 3 first strand kit, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

PerfeCTa SYBR Green FastMix green fluorescent dye PerfeCTa SYBR Green FastMix for iQ Quanta PerfeCta SYBR Green FastMix SYBR Green FastMix ROX PerfeCTa SYBR Green FastMix PerfeCTa SYBR Green FastMix Reaction Mixes PerfeCTa SYBR Green FastMix reagent PerfeCTa SYBR Green FastMix ROX PerfeCTa SYBR Green QPCR FastMix ROX Perfecta SYBR Green FastMix ROX kit

27 protocols using sybr green fastmix

1

ChIP-qPCR Analysis of Histone Modifications

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Chromatin was immunoprecipitated using antibodies against H3K27ac and H3K4me3, and gDNA was purified for qPCR analysis. All sequences for ChIP-qPCR primers can be found in Table S3. qPCR was performed using SYBR Green Fastmix (Quanta BioSciences), and the data are presented as fold change gDNA relative to negative control and normalized to a region of the Gapdh locus.
Black J.B., Adler A.F., Wang H.G., D’Ippolito A.M., Hutchinson H.A., Reddy T.E., Pitt G.S., Leong K.W, & Gersbach C.A. (2016). Targeted Epigenetic Remodeling of Endogenous Loci by CRISPR/Cas9-Based Transcriptional Activators Directly Converts Fibroblasts to Neuronal Cells. Cell stem cell, 19(3), 406-414.
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2

Quantifying Neuraminidase and Sialyltransferase Expression

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Total cellular RNA was isolated from N2a cells using Omega MicroElute Total RNA kit (Omega, Norcross, GA), according to the manufacturer's instructions. Subsequently, cDNA was synthesized from isolated RNA samples using the iScript cDNA Synthesis kit (Bio-Rad, Hercules, CA). Quantification of cDNA was done using a real-time RT-PCR machine (BioRad CFX96 Touch) and SYBR Green Fastmix (Quanta Biosciences, Gaithersburg, MD). Primers for detection of Neu1, Neu3, Neu, ST3Gal3, ST3Gal4, ST3Gal6, ST6Gal1 and ST6Gal2 were designed using Primer-BLAST and are reported in Table 2. The relative gene expression of neuraminidases and sialyltransferases was calculated using the ΔCt method, expression in four independent repeats was analyzed for each enzyme. The housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control for normalizing relative mRNA levels in all qRT-PCR experiments.
Katorcha E., Klimova N., Makarava N., Savtchenko R., Pan X., Annunziata I., Takahashi K., Miyagi T., Pshezhetsky A.V., d’Azzo A, & Baskakov I.V. (2015). Loss of Cellular Sialidases Does Not Affect the Sialylation Status of the Prion Protein but Increases the Amounts of Its Proteolytic Fragment C1. PLoS ONE, 10(11), e0143218.
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3

Extraction and Synthesis of RNA for qPCR

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Total RNA was purified using TRIzol Reagent (Thermo Fisher Scientific) following manufacturer's instructions and resuspended in RNAse-free water. cDNA was synthesized using high-capacity RNA-to-cDNA Kit (Applied Biosystems). SYBR Green FastMix (Quanta BioSciences) was used for the PCRs. Primer sequences were designed to span exon-exon junctions in Primer-BLAST (NCBI).
Calvo-Vidal M.N., Zamponi N., Krumsiek J., Stockslager M.A., Revuelta M.V., Phillip J.M., Marullo R., Tikhonova E., Kotlov N., Patel J., Yang S.N., Yang L., Taldone T., Thieblemont C., Leonard J.P., Martin P., Inghirami G., Chiosis G., Manalis S.R, & Cerchietti L. (2021). Oncogenic HSP90 Facilitates Metabolic Alterations in Aggressive B-cell Lymphomas. Cancer Research, 81(20), 5202-5216.
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4

Quantitative RT-PCR of Gene Expression

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Total RNA was isolated using TRIzol (Invitrogen), and was converted to cDNA using Superscript II reverse transcriptase (Invitrogen, USA). Quantitative RT-PCR (qRT-PCR) was performed with ABI Prism 7900 Sequence Detection System (Applied Biosystems, Foster City, CA) using SYBR Green FastMix (Quanta Biosciences, Gaithersburg, MD). GAPDH was used as a normalization gene and the standard curve method was used to calculate the relative expression. The PCR conditions used were 50°C for 2 min and 95°C for 10 min followed by 95°C for 15 s and 60°C for 40 s for 40 cycles. Primers used for qRT-PCR analysis are shown in S1 Table.
Dias H.B., de Oliveira J.R., Donadio M.V, & Kimura S. (2019). Fructose-1,6-bisphosphate prevents pulmonary fibrosis by regulating extracellular matrix deposition and inducing phenotype reversal of lung myofibroblasts. PLoS ONE, 14(9), e0222202.
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5

Silencing of Transcription Regulators

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ON-TARGETplus SMARTpool siRNAs targeting Arid5a, HuR, TRAF2 and MCPIP1 were from Dharmacon (Lafayette, CO). For RNA silencing, ST2 cells and MEFs were seeded overnight in antibiotic-free α-MEM, and N/TERT2G cells were seeded in KC-serum free media (Gibco) with supplements. Transfection was performed 18 h later with 50nM siRNAs with DharmaFECT Reagent 1. Culture media was replaced after 24 h, and IL-17 administered 24 h later. Flag/Myc- Arid5a was from OriGene (Rockville MD). All other constructs were described (6 (link), 22 (link), 27 (link), 63 (link)). Transfections of ST2 cells and MEFS were performed with Fugene HD (Promega, MadisonWI) or CaPO4 (HEK293T cells). RNA was prepared with RNeasy Kits (Qiagen, Valencia CA). cDNA was synthesized by Superscript III First Strand Kits (ThermoFisher, Waltham MA). Quantitative real-time PCR (qPCR) was performed with the SYBR Green FastMix (Quanta Biosciences, Beverly MA) and analyzed on a 7300 ABI Real Time instrument. Primers were from QuantiTect Primer Assays (Qiagen).
Amatya N., Childs E.E., Cruz J.A., Aggor F.E., Garg A.V., Berman A.J., Gudjonsson J.E., Atasoy U, & Gaffen S.L. (2018). IL-17 integrates multiple self-reinforcing, feed-forward mechanisms through the RNA-binding protein Arid5a. Science signaling, 11(551), eaat4617.
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6

Quantitative Real-Time PCR Analysis of Angiogenin

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Total RNA was extracted using RNAzol RT (Sigma-Aldrich) and quantified using a Nanodrop 2000 Spectrophotometer (ThermoScientific). A total of 600 ng RNA was reverse transcribed (qScript™ cDNA Supermix, Quanta Biosciences, Beverly, MA, USA) to obtain cDNA (25 °C × 5 min, 42 °C × 30 min, 85 °C × 5 min, and hold at 4 °C). For quantitative real-time PCR, human angiogenin primers (Hs ANG 2 SG) (Qiagene, Toronto, ON, Canada, Cat# QT01675212) and GAPDH primers (Qiagene, Cat#QT01192646) were used with SYBR Green Fast mix (Quanta Bioscience) and samples were tested in triplicate with the following cycling conditions: 95 °C × 2 min, 95 °C × 10 s, 55 °C × 30 s, and 72 °C × 10 s for 40 cycles (Mastercycler® ep realplex, Eppendorf, Mississauga, ON, Canada). Primer efficiencies were tested, and standard curves were generated for each primer set.
Fatica T., Naas T., Liwak U., Slaa H., Souaid M., Frangione B., Kattini R., Gaudreau-Lapierre A., Trinkle-Mulcahy L., Chakraborty P, & Holcik M. (2023). TRNT-1 Deficiency Is Associated with Loss of tRNA Integrity and Imbalance of Distinct Proteins. Genes, 14(5), 1043.
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7

RNA Isolation and qPCR Analysis

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RNA was isolated by extraction with Trizol according to manufacturer instructions (Invitrogen) or with the RNA Clean & Concentrator Kit (Zymo Research). cDNA was synthesized using iScript cDNA synthesis kit (BioRad). Quantitative real-time PCR was performed with SYBR Green Fast Mix (Quanta Biosciences) on a Roche Lightcycler 480 and analyzed by using ΔΔCt calculations. qPCR analyses in human cell lines are relative to the reference gene B2M. qPCR analyses in mouse cell line or tissue are relative to β-Actin. The primer sequences are described in Table S1.
Yoon H., Spinelli J.B., Zaganjor E., Wong S.J., German N.J., Randall E.C., Dean A., Clermont A., Paulo J.A., Garcia D., Li H., Rombold O., Agar N.Y., Goodyear L.J., Shaw R.J., Gygi S.P., Auwerx J, & Haigis M.C. (2020). PHD3 loss promotes exercise capacity and fat oxidation in skeletal muscle. Cell metabolism, 32(2), 215-228.e7.
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8

Circadian Regulation of Per2 Gene Expression

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Mice were sacrificed by sodium pentobarbital injection immediately prior to tissue collection at ZT 18, ZT 0, ZT 6, or ZT 12. Tissue was rapidly extracted, frozen on dry ice, and stored at −80°C until RNA extraction. Tissue samples were later mechanically homogenized and RNA was extracted using Trizol Reagent (Invitrogen) according to manufacturer's specifications. RNA concentrations were obtained using a Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA) and concentrations were adjusted to 50 ng/µl. cDNA was synthesized from 500 ng RNA using High Capacity Reverse Transcription Kit (Life Technologies). Real-time PCR was performed using 2 µl cDNA with SYBR Green FastMix (Quanta Biosciences, Gaithersburg, MD) in a Step-One real time PCR system (Life Technologies). Gene expression was normalized to total RNA input. Primers used for PCR were as follows: Per2 forward: ACCTCCCTGCAGACAAGAA, Per2 reverse: CTCATTAGCCTTCACCTGCTT.
Gallardo C.M., Darvas M., Oviatt M., Chang C.H., Michalik M., Huddy T.F., Meyer E.E., Shuster S.A., Aguayo A., Hill E.M., Kiani K., Ikpeazu J., Martinez J.S., Purpura M., Smit A.N., Patton D.F., Mistlberger R.E., Palmiter R.D, & Steele A.D. (2014). Dopamine receptor 1 neurons in the dorsal striatum regulate food anticipatory circadian activity rhythms in mice. eLife, 3, e03781.
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9

Lung Total RNA Extraction and qPCR

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Total RNA was isolated from the whole lung according to the TRIzol reagent protocol (Invitrogen, NY, USA) and purified by E.Z.N.A. total RNA kit I (OMEGA, GA, USA). Reverse transcription PCR was performed using the qScript cDNA SuperMix (Quanta Biosciences, Gaithersburg, MD). qPCR was performed according to a standard protocol using SYBR Green FastMix, Low ROX (Quanta Biosciences, Gaithersburg, MD) and products measured on an ABI Viia 7 PCR system (ABI, Foster City, CA). Data are presented as fold change between the test groups and controls, as indicated in the figure legends. Gene-specific primers are listed in Supplementary Table 4.
Caetano M.S., Hassane M., Van H.T., Bugarin E., Cumpian A.M., McDowell C.L., Cavazos C.G., Zhang H., Deng S., Diao L., Wang J., Evans S.E., Behrens C., Wistuba I.I., Fuqua S.A., Lin H., Stabile L.P., Watowich S.S., Kadara H, & Moghaddam S.J. (2018). Sex specific function of epithelial STAT3 signaling in pathogenesis of K-ras mutant lung cancer. Nature Communications, 9, 4589.
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10

RNA Extraction and qPCR Analysis

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Total RNA was purified using TRIzol Reagent (Thermo Fisher Scientific) following manufacturer's instructions and resuspended in RNAse-free water. cDNA was synthesized using high capacity RNA-to-cDNA kit (Applied Biosystems). SYBR Green FastMix was from Quanta BioSciences. Primer sequences can be found in Table S4.
Pera B., Krumsiek J., Assouline S.E., Marullo R., Patel J., Phillip J.M., Román L., Mann K.K, & Cerchietti L. (2018). Metabolomic Profiling Reveals Cellular Reprogramming of B-Cell Lymphoma by a Lysine Deacetylase Inhibitor through the Choline Pathway. EBioMedicine, 28, 80-89.
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