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TaKaRa sybr premix ex taq ii
Quantitative analysis of NOX2 and NOX4 mRNA expression in human OS cell lines. Quantitative analysis of the mRNA expression of NOX family proteins in five human OS cell lines was performed by RT-qPCR using a <t>SYBR</t> Green reagent. The Cq value during the exponential phase of amplification was determined by the real-time monitoring of the fluorescent emission of <t>Taq</t> polymerase nuclease activity. β- actin was used as an internal control for normalization. qPCR efficiencies of the target ( NOX2 and NOX4 ) and control (β-actin) proteins were approximately equal over a concentration range of 0.1–200 ng total cDNA. The relative transcripts were determined by the formula: 1/2 (Cq target-Cq control) ). The bars represent the mean (n=2). OS, osteosarcoma; NOX, NADPH oxidase; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; Cq, quantification cycle; PBMCs, peripheral blood mononuclear cells.
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1) Product Images from "Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation"

Article Title: Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation

Journal: Oncology Letters

doi: 10.3892/ol.2018.8291

Quantitative analysis of NOX2 and NOX4 mRNA expression in human OS cell lines. Quantitative analysis of the mRNA expression of NOX family proteins in five human OS cell lines was performed by RT-qPCR using a SYBR Green reagent. The Cq value during the exponential phase of amplification was determined by the real-time monitoring of the fluorescent emission of Taq polymerase nuclease activity. β- actin was used as an internal control for normalization. qPCR efficiencies of the target ( NOX2 and NOX4 ) and control (β-actin) proteins were approximately equal over a concentration range of 0.1–200 ng total cDNA. The relative transcripts were determined by the formula: 1/2 (Cq target-Cq control) ). The bars represent the mean (n=2). OS, osteosarcoma; NOX, NADPH oxidase; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; Cq, quantification cycle; PBMCs, peripheral blood mononuclear cells.
Figure Legend Snippet: Quantitative analysis of NOX2 and NOX4 mRNA expression in human OS cell lines. Quantitative analysis of the mRNA expression of NOX family proteins in five human OS cell lines was performed by RT-qPCR using a SYBR Green reagent. The Cq value during the exponential phase of amplification was determined by the real-time monitoring of the fluorescent emission of Taq polymerase nuclease activity. β- actin was used as an internal control for normalization. qPCR efficiencies of the target ( NOX2 and NOX4 ) and control (β-actin) proteins were approximately equal over a concentration range of 0.1–200 ng total cDNA. The relative transcripts were determined by the formula: 1/2 (Cq target-Cq control) ). The bars represent the mean (n=2). OS, osteosarcoma; NOX, NADPH oxidase; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; Cq, quantification cycle; PBMCs, peripheral blood mononuclear cells.

Techniques Used: Expressing, Quantitative RT-PCR, SYBR Green Assay, Amplification, Activity Assay, Real-time Polymerase Chain Reaction, Concentration Assay

Related Articles

Amplification:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: The specific primers (Bioneer, Daejeon, South Korea) for measurement of target genes and GAPDH were used and the lengths for amplified products were as follow: GAPDH (5´-TGCTGGTGCTGAGTATGTCGTG-3 and 5´ CGG AGATGA TGACCCTTTTGG-3´, 101 bp); NBC1(5´-CTCACAT TCGGTCGCCAAAC-3´and 5´-CCCCAATGTAGATC GTGT GGT-3, 149 bp); TFF1 (5´-TCCCTTCTAAGGTCCAT CCGA-3´and 5´-CAGATCCA GCCACCGAACAG-3, 126 bp); and TFF2 (5´-GACGC CCTCCAACAG AAAGA-3´and 5´CGACGCTT GGTTTGGAAGT G-3, 133 bp). .. All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min.

Article Title: The Fungal and Bacterial Rhizosphere Microbiome Associated With Grapevine Rootstock Genotypes in Mature and Young Vineyards
Article Snippet: Paragraph title: Quantitative PCR Amplification and Quantification of Black-Foot Disease Pathogens ... Each reaction contained 2 μl of DNA, 1× of SYBR Premix Ex Taq II (Tli RNase H Plus) (Takara Bio Inc., Shiga, Japan) and 0.4 μM of each primer.

Article Title: ∆133p53 isoform promotes tumour invasion and metastasis via interleukin-6 activation of JAK-STAT and RhoA-ROCK signalling
Article Snippet: RT-qPCR was performed as follows: 100 ng of cDNA was added to 10 µl of SYBR Premix Ex Taq II (Ti RNase H Plus; Takara Bio, Japan) and 200 nM of each primer, in a final volume of 20 µl; the PCR product was run on the Roche LightCycler 480 as follows: 95 °C for 2 min, then 40 cycles of 95 °C for 30 s, 60 °C for 1 min, followed by a melt curve analysis; and RT-qPCR was performed for each sample with each primer pair in triplicate. .. Relative transcript abundance from RT-qPCR was calculated using the equation: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\rm Amplification\,efficiency}^{ \wedge - ({\mathrm{geometric}}\,{\mathrm{mean}}\,{\mathrm{reference}}\,{\mathrm{gene}}\,{\mathrm{threshold}}\,{\mathrm{Ct}}\,-TP53\ {\mathrm{threshold}}\,{\mathrm{Ct}})}$$\end{document} A m p l i f i c a t i o n e f f i c i e n c y ∧ - ( geometric mean reference gene threshold Ct - T P 53 threshold Ct )

Positive Control:

Article Title: The Fungal and Bacterial Rhizosphere Microbiome Associated With Grapevine Rootstock Genotypes in Mature and Young Vineyards
Article Snippet: Each reaction contained 2 μl of DNA, 1× of SYBR Premix Ex Taq II (Tli RNase H Plus) (Takara Bio Inc., Shiga, Japan) and 0.4 μM of each primer. .. Each analysis included three replicates of each sample, a non-template control reaction (water) and a positive control containing DNA extracted from a pure culture of the Dactylonectria torresensis isolate GTMF DT097, obtained from the collection of the Instituto Agroforestal Mediterráneo, Universitat Politècnica de Valencia, Spain.

Synthesized:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: The cDNA was synthesized from one microgram of the total RNA using QuantiTect reverse transcription Kit (Qiagen) according to the manufacturer’s instruction. .. All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min.

Article Title: Plant A20/AN1 protein serves as the important hub to mediate antiviral immunity
Article Snippet: For qRT-PCR, cDNA was synthesized from 500 ng of DNA-free RNA and oligo (dT) by use of PrimeScript RT Reagent Kit (Perfect Real Time) (Takara Bio) following the manufacturer’s instructions (Takara Bio). .. The cDNA template was used for qPCR by use of SYBR Premix EX Taq II (Ti RNase H Plus) Kit (Takara Bio) in an ABI Prism 7500 sequence detection system (Applied Biosystems).

Article Title: Omeprazole Inhibits Cell Proliferation and Induces G0/G1 Cell Cycle Arrest through Up-regulating miR-203a-3p Expression in Barrett’s Esophagus Cells
Article Snippet: .. For mRNA analysis, the first-strand cDNA was synthesized using a PrimeScript RT Reagent Kit (Takara, Japan) in accordance with the protocol of manufacturer. qRT-PCR was used for the detection of target gene expression by employing the SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States). miRNA expression was determined with qRT-PCR, to this end, the first-strand cDNA was synthesized using Mir-XTM miRNA First-Strand Synthesis Kit (Takara, Japan) and qRT-PCR was conducted with SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States). ..

Article Title: Transcriptome Analysis of the Breast Muscle of Xichuan Black-Bone Chickens Under Tyrosine Supplementation Revealed the Mechanism of Tyrosine-Induced Melanin Deposition
Article Snippet: The gene-specific primers were designed using the NCBI Primer BLAST online program ( ) and synthesized by Nanjing Kingsray Biotechnology Co., Ltd. .. The qRT-PCR analysis was performed using a PrimeScriptTM RT Reagent Kit and SYBR® Premix Ex Taq II (Tli RNase H Plus; Takara, Dalian, China).

Quantitative RT-PCR:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: The mRNA levels of the NBC1, TFF1, TFF2, and the housekeeping gene glycer-aldehyde-3-phosphate dehydrogenase (GAPDH) were measured by quantitative real-time PCR (qRT-PCR) using LightCycler® 480 system (Roche Diagnostics, Branford, USA). .. All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min.

Article Title: Plant A20/AN1 protein serves as the important hub to mediate antiviral immunity
Article Snippet: Paragraph title: RNA isolation, semi-quantitative and qRT-PCR analysis ... The cDNA template was used for qPCR by use of SYBR Premix EX Taq II (Ti RNase H Plus) Kit (Takara Bio) in an ABI Prism 7500 sequence detection system (Applied Biosystems).

Article Title: Omeprazole Inhibits Cell Proliferation and Induces G0/G1 Cell Cycle Arrest through Up-regulating miR-203a-3p Expression in Barrett’s Esophagus Cells
Article Snippet: .. For mRNA analysis, the first-strand cDNA was synthesized using a PrimeScript RT Reagent Kit (Takara, Japan) in accordance with the protocol of manufacturer. qRT-PCR was used for the detection of target gene expression by employing the SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States). miRNA expression was determined with qRT-PCR, to this end, the first-strand cDNA was synthesized using Mir-XTM miRNA First-Strand Synthesis Kit (Takara, Japan) and qRT-PCR was conducted with SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States). ..

Article Title: Site-specific Proteasome Phosphorylation Controls Cell Proliferation and Tumorigenesis
Article Snippet: Paragraph title: Quantitative RT-PCR ... Gene-specific primers and cDNAs were mixed with SYBR Premix Ex Taq II (Tli RNase H Plus, Takara) for PCR reactions in a 7500 Real-Time PCR System (Applied Biosystems).

Article Title: Expression of antioxidant genes in broiler chickens fed nettle (Urtica dioica) and its link with pulmonary hypertension
Article Snippet: .. The levels of superoxide dismutase 1 (SOD1 ), catalase (CAT ), glutathione peroxidase (GPX ) and β-actin transcripts were determined by quantitative real time (RT)-PCR using SYBR Premix Ex Taq II (Tli Rnase H Plus) (TaKaRa Bio Inc., Japan). ..

Article Title: Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation
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Article Title: Transcriptome Analysis of the Breast Muscle of Xichuan Black-Bone Chickens Under Tyrosine Supplementation Revealed the Mechanism of Tyrosine-Induced Melanin Deposition
Article Snippet: .. The qRT-PCR analysis was performed using a PrimeScriptTM RT Reagent Kit and SYBR® Premix Ex Taq II (Tli RNase H Plus; Takara, Dalian, China). ..

Article Title: ∆133p53 isoform promotes tumour invasion and metastasis via interleukin-6 activation of JAK-STAT and RhoA-ROCK signalling
Article Snippet: .. RT-qPCR was performed as follows: 100 ng of cDNA was added to 10 µl of SYBR Premix Ex Taq II (Ti RNase H Plus; Takara Bio, Japan) and 200 nM of each primer, in a final volume of 20 µl; the PCR product was run on the Roche LightCycler 480 as follows: 95 °C for 2 min, then 40 cycles of 95 °C for 30 s, 60 °C for 1 min, followed by a melt curve analysis; and RT-qPCR was performed for each sample with each primer pair in triplicate. .. Relative transcript abundance from RT-qPCR was calculated using the equation: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\rm Amplification\,efficiency}^{ \wedge - ({\mathrm{geometric}}\,{\mathrm{mean}}\,{\mathrm{reference}}\,{\mathrm{gene}}\,{\mathrm{threshold}}\,{\mathrm{Ct}}\,-TP53\ {\mathrm{threshold}}\,{\mathrm{Ct}})}$$\end{document} A m p l i f i c a t i o n e f f i c i e n c y ∧ - ( geometric mean reference gene threshold Ct - T P 53 threshold Ct )

SYBR Green Assay:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: .. All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min. .. In addition, the no-template negative control (H2 O) was routinely run in every examined at the end of amplification process to ensure the specificity of PCR products.

Article Title: Redirecting abiraterone metabolism to fine tune prostate cancer anti-androgen therapy
Article Snippet: Quantitative PCR (qPCR) analysis was conducted in triplicate in an ABI 7500 Real-Time PCR machine (Applied Biosystems) using iTaq Fast SYBR Green Supermix with ROX (Bio-Rad) and primers for TMPRSS2 , PSA and RPLPO , as described previously . .. SYBR Premix Ex Taq II (Takara) was used for SRD5A1 , SRD5A2 and AR v7 detection.

Incubation:

Article Title: Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation
Article Snippet: RT was conducted as follows: A total of 8 µl water, containing 1 µg total RNA, was added to 50 ng random primers (Thermo-Fisher Scientific, Inc.) and incubated at 65°C for 5 min. .. RT-qPCR was performed with SYBR Premix Ex Taq II (Takara Bio, Inc., Otsu, Shiga, Japan) in an ABI PRISM 7500 Sequence Detection system (Applied Biosystems; Thermo Fisher Scientific, Inc.).

Expressing:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min. .. Expression level of NBC1, TFF1 and TFF2 were normalized against GAPDH expression (internal calibrator for equal RNA template loading and normalization).

Article Title: Spinal cord reconstitution with homologous neural grafts enables robust corticospinal regeneration
Article Snippet: For cDNA synthesis, the reverse transcription reaction was performed using the PrimeScript™ RT Master Mix (Perfect Real Time, Clonthech) and quantitative PCR was performed using primers listed in and SYBR® Premix Ex Taq™ II (Tli RNase H Plus, Clontech) in 20-μL reactions. .. Data were normalized to glyceraldehydes-3-phosphate dehydrogenase expression (GAPDH).

Article Title: Omeprazole Inhibits Cell Proliferation and Induces G0/G1 Cell Cycle Arrest through Up-regulating miR-203a-3p Expression in Barrett’s Esophagus Cells
Article Snippet: .. For mRNA analysis, the first-strand cDNA was synthesized using a PrimeScript RT Reagent Kit (Takara, Japan) in accordance with the protocol of manufacturer. qRT-PCR was used for the detection of target gene expression by employing the SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States). miRNA expression was determined with qRT-PCR, to this end, the first-strand cDNA was synthesized using Mir-XTM miRNA First-Strand Synthesis Kit (Takara, Japan) and qRT-PCR was conducted with SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States). ..

Article Title: Redirecting abiraterone metabolism to fine tune prostate cancer anti-androgen therapy
Article Snippet: Paragraph title: Gene expression and immunoblotting ... SYBR Premix Ex Taq II (Takara) was used for SRD5A1 , SRD5A2 and AR v7 detection.

Article Title: Transcriptome Analysis of the Breast Muscle of Xichuan Black-Bone Chickens Under Tyrosine Supplementation Revealed the Mechanism of Tyrosine-Induced Melanin Deposition
Article Snippet: The qRT-PCR analysis was performed using a PrimeScriptTM RT Reagent Kit and SYBR® Premix Ex Taq II (Tli RNase H Plus; Takara, Dalian, China). .. The relative expression levels of the genes were calculated using the 2-ΔΔCt method, and the levels were normalized to those of two housekeeping genes, β-actin and GAPDH ( ).

Reverse Transcription Polymerase Chain Reaction:

Article Title: Spinal cord reconstitution with homologous neural grafts enables robust corticospinal regeneration
Article Snippet: Paragraph title: Quantitative reverse transcription-polymerase chain reaction (RT-PCR) ... For cDNA synthesis, the reverse transcription reaction was performed using the PrimeScript™ RT Master Mix (Perfect Real Time, Clonthech) and quantitative PCR was performed using primers listed in and SYBR® Premix Ex Taq™ II (Tli RNase H Plus, Clontech) in 20-μL reactions.

Article Title: Plant A20/AN1 protein serves as the important hub to mediate antiviral immunity
Article Snippet: The results of semi-quantitative RT-PCR were analyzed by ImageJ software for the relative quantification. .. The cDNA template was used for qPCR by use of SYBR Premix EX Taq II (Ti RNase H Plus) Kit (Takara Bio) in an ABI Prism 7500 sequence detection system (Applied Biosystems).

Article Title: Expression of antioxidant genes in broiler chickens fed nettle (Urtica dioica) and its link with pulmonary hypertension
Article Snippet: The levels of superoxide dismutase 1 (SOD1 ), catalase (CAT ), glutathione peroxidase (GPX ) and β-actin transcripts were determined by quantitative real time (RT)-PCR using SYBR Premix Ex Taq II (Tli Rnase H Plus) (TaKaRa Bio Inc., Japan). .. PCR were carried out in a RT PCR cycler (Rotor Gene Q 6000, Qiagen, USA) in 3 replicates for each sample of ventricles.

Negative Control:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min. .. In addition, the no-template negative control (H2 O) was routinely run in every examined at the end of amplification process to ensure the specificity of PCR products.

Sequencing:

Article Title: Plant A20/AN1 protein serves as the important hub to mediate antiviral immunity
Article Snippet: .. The cDNA template was used for qPCR by use of SYBR Premix EX Taq II (Ti RNase H Plus) Kit (Takara Bio) in an ABI Prism 7500 sequence detection system (Applied Biosystems). ..

Article Title: Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation
Article Snippet: .. RT-qPCR was performed with SYBR Premix Ex Taq II (Takara Bio, Inc., Otsu, Shiga, Japan) in an ABI PRISM 7500 Sequence Detection system (Applied Biosystems; Thermo Fisher Scientific, Inc.). .. Briefly, a solution of SYBR Premix Ex Taq II (10 µl) containing sense and antisense primers (10 µM each) was prepared and aliquoted into individual wells of a MicroAmp Optical Plate (ABI-PE; Applied Biosystems; Thermo-Fisher Scientific, Inc.): 2 µl cDNA was added to give a final volume of 20 µl.

RNA Sequencing Assay:

Article Title: Transcriptome Analysis of the Breast Muscle of Xichuan Black-Bone Chickens Under Tyrosine Supplementation Revealed the Mechanism of Tyrosine-Induced Melanin Deposition
Article Snippet: Quantitative Real-Time PCR (qRT-PCR) Analysis To verify the accuracy of the RNA-seq results, seven genes were selected for qRT-PCR validation. .. The qRT-PCR analysis was performed using a PrimeScriptTM RT Reagent Kit and SYBR® Premix Ex Taq II (Tli RNase H Plus; Takara, Dalian, China).

Isolation:

Article Title: Spinal cord reconstitution with homologous neural grafts enables robust corticospinal regeneration
Article Snippet: Quantitative reverse transcription-polymerase chain reaction (RT-PCR) Total RNA was isolated from neural progenitor cell cultures using the RNeasy Mini kit (Qiagen, Germantown, PA) following the manufacturer’s protocol. .. For cDNA synthesis, the reverse transcription reaction was performed using the PrimeScript™ RT Master Mix (Perfect Real Time, Clonthech) and quantitative PCR was performed using primers listed in and SYBR® Premix Ex Taq™ II (Tli RNase H Plus, Clontech) in 20-μL reactions.

Article Title: Plant A20/AN1 protein serves as the important hub to mediate antiviral immunity
Article Snippet: Paragraph title: RNA isolation, semi-quantitative and qRT-PCR analysis ... The cDNA template was used for qPCR by use of SYBR Premix EX Taq II (Ti RNase H Plus) Kit (Takara Bio) in an ABI Prism 7500 sequence detection system (Applied Biosystems).

Article Title: ∆133p53 isoform promotes tumour invasion and metastasis via interleukin-6 activation of JAK-STAT and RhoA-ROCK signalling
Article Snippet: RNA extraction and RT-qPCR Total RNA was isolated from snap-frozen tumours as per the manufacturer’s protocol using the PureLink RNA mini kit (Invitrogen, USA). .. RT-qPCR was performed as follows: 100 ng of cDNA was added to 10 µl of SYBR Premix Ex Taq II (Ti RNase H Plus; Takara Bio, Japan) and 200 nM of each primer, in a final volume of 20 µl; the PCR product was run on the Roche LightCycler 480 as follows: 95 °C for 2 min, then 40 cycles of 95 °C for 30 s, 60 °C for 1 min, followed by a melt curve analysis; and RT-qPCR was performed for each sample with each primer pair in triplicate.

Size-exclusion Chromatography:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: .. All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min. .. In addition, the no-template negative control (H2 O) was routinely run in every examined at the end of amplification process to ensure the specificity of PCR products.

Article Title: Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation
Article Snippet: RT-qPCR was performed with SYBR Premix Ex Taq II (Takara Bio, Inc., Otsu, Shiga, Japan) in an ABI PRISM 7500 Sequence Detection system (Applied Biosystems; Thermo Fisher Scientific, Inc.). .. The cycling conditions for the PCR were 42°C for 5 min, 95°C for 10 sec and 40 cycles of 95°C for 5 sec (denaturation) and 60°C for 34 sec (annealing/extension).

Purification:

Article Title: Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation
Article Snippet: Purified RNA was dissolved in 20 µl distilled water. .. RT-qPCR was performed with SYBR Premix Ex Taq II (Takara Bio, Inc., Otsu, Shiga, Japan) in an ABI PRISM 7500 Sequence Detection system (Applied Biosystems; Thermo Fisher Scientific, Inc.).

Polymerase Chain Reaction:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: .. All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min. .. In addition, the no-template negative control (H2 O) was routinely run in every examined at the end of amplification process to ensure the specificity of PCR products.

Article Title: Plant A20/AN1 protein serves as the important hub to mediate antiviral immunity
Article Snippet: The PCR was performed using the gene-specific primer pairs ( ). .. The cDNA template was used for qPCR by use of SYBR Premix EX Taq II (Ti RNase H Plus) Kit (Takara Bio) in an ABI Prism 7500 sequence detection system (Applied Biosystems).

Article Title: Site-specific Proteasome Phosphorylation Controls Cell Proliferation and Tumorigenesis
Article Snippet: .. Gene-specific primers and cDNAs were mixed with SYBR Premix Ex Taq II (Tli RNase H Plus, Takara) for PCR reactions in a 7500 Real-Time PCR System (Applied Biosystems). ..

Article Title: Expression of antioxidant genes in broiler chickens fed nettle (Urtica dioica) and its link with pulmonary hypertension
Article Snippet: The levels of superoxide dismutase 1 (SOD1 ), catalase (CAT ), glutathione peroxidase (GPX ) and β-actin transcripts were determined by quantitative real time (RT)-PCR using SYBR Premix Ex Taq II (Tli Rnase H Plus) (TaKaRa Bio Inc., Japan). .. PCR were carried out in a RT PCR cycler (Rotor Gene Q 6000, Qiagen, USA) in 3 replicates for each sample of ventricles.

Article Title: Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation
Article Snippet: Paragraph title: Quantification of NOX2 and NOX4 mRNAs by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) ... RT-qPCR was performed with SYBR Premix Ex Taq II (Takara Bio, Inc., Otsu, Shiga, Japan) in an ABI PRISM 7500 Sequence Detection system (Applied Biosystems; Thermo Fisher Scientific, Inc.).

Article Title: ∆133p53 isoform promotes tumour invasion and metastasis via interleukin-6 activation of JAK-STAT and RhoA-ROCK signalling
Article Snippet: .. RT-qPCR was performed as follows: 100 ng of cDNA was added to 10 µl of SYBR Premix Ex Taq II (Ti RNase H Plus; Takara Bio, Japan) and 200 nM of each primer, in a final volume of 20 µl; the PCR product was run on the Roche LightCycler 480 as follows: 95 °C for 2 min, then 40 cycles of 95 °C for 30 s, 60 °C for 1 min, followed by a melt curve analysis; and RT-qPCR was performed for each sample with each primer pair in triplicate. .. Relative transcript abundance from RT-qPCR was calculated using the equation: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\rm Amplification\,efficiency}^{ \wedge - ({\mathrm{geometric}}\,{\mathrm{mean}}\,{\mathrm{reference}}\,{\mathrm{gene}}\,{\mathrm{threshold}}\,{\mathrm{Ct}}\,-TP53\ {\mathrm{threshold}}\,{\mathrm{Ct}})}$$\end{document} A m p l i f i c a t i o n e f f i c i e n c y ∧ - ( geometric mean reference gene threshold Ct - T P 53 threshold Ct )

Chloramphenicol Acetyltransferase Assay:

Article Title: Expression of antioxidant genes in broiler chickens fed nettle (Urtica dioica) and its link with pulmonary hypertension
Article Snippet: .. The levels of superoxide dismutase 1 (SOD1 ), catalase (CAT ), glutathione peroxidase (GPX ) and β-actin transcripts were determined by quantitative real time (RT)-PCR using SYBR Premix Ex Taq II (Tli Rnase H Plus) (TaKaRa Bio Inc., Japan). ..

Plasmid Preparation:

Article Title: Bacterial Communities Changes during Food Waste Spoilage
Article Snippet: Quantitative PCR was performed in a 25-μL reaction mixture containing 12.5 μL SYBR Premix Ex Taq II (Tli RNase H Plus, 2×; Takara), 0.48 μM each primer, and 2 μL template DNA. .. The standard template plasmid DNA was diluted in a 10−1 to 1−8 series using EASY Dilution for Real Time PCR (Takara).

Software:

Article Title: Plant A20/AN1 protein serves as the important hub to mediate antiviral immunity
Article Snippet: The results of semi-quantitative RT-PCR were analyzed by ImageJ software for the relative quantification. .. The cDNA template was used for qPCR by use of SYBR Premix EX Taq II (Ti RNase H Plus) Kit (Takara Bio) in an ABI Prism 7500 sequence detection system (Applied Biosystems).

Article Title: Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation
Article Snippet: RT-qPCR was performed with SYBR Premix Ex Taq II (Takara Bio, Inc., Otsu, Shiga, Japan) in an ABI PRISM 7500 Sequence Detection system (Applied Biosystems; Thermo Fisher Scientific, Inc.). .. The data were analyzed with Sequence Detector software (version 1.6; ABI-PE; Applied Biosystems; Thermo-Fisher Scientific, Inc.).

Real-time Polymerase Chain Reaction:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: The mRNA levels of the NBC1, TFF1, TFF2, and the housekeeping gene glycer-aldehyde-3-phosphate dehydrogenase (GAPDH) were measured by quantitative real-time PCR (qRT-PCR) using LightCycler® 480 system (Roche Diagnostics, Branford, USA). .. All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min.

Article Title: Spinal cord reconstitution with homologous neural grafts enables robust corticospinal regeneration
Article Snippet: .. For cDNA synthesis, the reverse transcription reaction was performed using the PrimeScript™ RT Master Mix (Perfect Real Time, Clonthech) and quantitative PCR was performed using primers listed in and SYBR® Premix Ex Taq™ II (Tli RNase H Plus, Clontech) in 20-μL reactions. .. Data were normalized to glyceraldehydes-3-phosphate dehydrogenase expression (GAPDH).

Article Title: Plant A20/AN1 protein serves as the important hub to mediate antiviral immunity
Article Snippet: .. The cDNA template was used for qPCR by use of SYBR Premix EX Taq II (Ti RNase H Plus) Kit (Takara Bio) in an ABI Prism 7500 sequence detection system (Applied Biosystems). ..

Article Title: Omeprazole Inhibits Cell Proliferation and Induces G0/G1 Cell Cycle Arrest through Up-regulating miR-203a-3p Expression in Barrett’s Esophagus Cells
Article Snippet: .. For mRNA analysis, the first-strand cDNA was synthesized using a PrimeScript RT Reagent Kit (Takara, Japan) in accordance with the protocol of manufacturer. qRT-PCR was used for the detection of target gene expression by employing the SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States). miRNA expression was determined with qRT-PCR, to this end, the first-strand cDNA was synthesized using Mir-XTM miRNA First-Strand Synthesis Kit (Takara, Japan) and qRT-PCR was conducted with SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States). ..

Article Title: Site-specific Proteasome Phosphorylation Controls Cell Proliferation and Tumorigenesis
Article Snippet: .. Gene-specific primers and cDNAs were mixed with SYBR Premix Ex Taq II (Tli RNase H Plus, Takara) for PCR reactions in a 7500 Real-Time PCR System (Applied Biosystems). ..

Article Title: Expression of antioxidant genes in broiler chickens fed nettle (Urtica dioica) and its link with pulmonary hypertension
Article Snippet: Paragraph title: Quantitative real time PCR analysis ... The levels of superoxide dismutase 1 (SOD1 ), catalase (CAT ), glutathione peroxidase (GPX ) and β-actin transcripts were determined by quantitative real time (RT)-PCR using SYBR Premix Ex Taq II (Tli Rnase H Plus) (TaKaRa Bio Inc., Japan).

Article Title: The Fungal and Bacterial Rhizosphere Microbiome Associated With Grapevine Rootstock Genotypes in Mature and Young Vineyards
Article Snippet: Paragraph title: Quantitative PCR Amplification and Quantification of Black-Foot Disease Pathogens ... Each reaction contained 2 μl of DNA, 1× of SYBR Premix Ex Taq II (Tli RNase H Plus) (Takara Bio Inc., Shiga, Japan) and 0.4 μM of each primer.

Article Title: Redirecting abiraterone metabolism to fine tune prostate cancer anti-androgen therapy
Article Snippet: Quantitative PCR (qPCR) analysis was conducted in triplicate in an ABI 7500 Real-Time PCR machine (Applied Biosystems) using iTaq Fast SYBR Green Supermix with ROX (Bio-Rad) and primers for TMPRSS2 , PSA and RPLPO , as described previously . .. SYBR Premix Ex Taq II (Takara) was used for SRD5A1 , SRD5A2 and AR v7 detection.

Article Title: Transcriptome Analysis of the Breast Muscle of Xichuan Black-Bone Chickens Under Tyrosine Supplementation Revealed the Mechanism of Tyrosine-Induced Melanin Deposition
Article Snippet: Paragraph title: Quantitative Real-Time PCR (qRT-PCR) Analysis ... The qRT-PCR analysis was performed using a PrimeScriptTM RT Reagent Kit and SYBR® Premix Ex Taq II (Tli RNase H Plus; Takara, Dalian, China).

Article Title: Bacterial Communities Changes during Food Waste Spoilage
Article Snippet: .. Quantitative PCR was performed in a 25-μL reaction mixture containing 12.5 μL SYBR Premix Ex Taq II (Tli RNase H Plus, 2×; Takara), 0.48 μM each primer, and 2 μL template DNA. ..

RNA Extraction:

Article Title: Omeprazole Inhibits Cell Proliferation and Induces G0/G1 Cell Cycle Arrest through Up-regulating miR-203a-3p Expression in Barrett’s Esophagus Cells
Article Snippet: Paragraph title: Total RNA Extraction and Quantitative Real-Time PCR (qRT-PCR) ... For mRNA analysis, the first-strand cDNA was synthesized using a PrimeScript RT Reagent Kit (Takara, Japan) in accordance with the protocol of manufacturer. qRT-PCR was used for the detection of target gene expression by employing the SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States). miRNA expression was determined with qRT-PCR, to this end, the first-strand cDNA was synthesized using Mir-XTM miRNA First-Strand Synthesis Kit (Takara, Japan) and qRT-PCR was conducted with SYBR Premix Ex Taq II (Takara, Japan) on an ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Grand Island, NY, United States).

Article Title: Redirecting abiraterone metabolism to fine tune prostate cancer anti-androgen therapy
Article Snippet: RNA extraction and cDNA synthesis were performed with the GenElute Mammalian Total RNA miniprep kit (Sigma-Aldrich) and iScript cDNA Synthesis Kit (Bio-Rad, Hercules, CA) respectively. .. SYBR Premix Ex Taq II (Takara) was used for SRD5A1 , SRD5A2 and AR v7 detection.

Article Title: ∆133p53 isoform promotes tumour invasion and metastasis via interleukin-6 activation of JAK-STAT and RhoA-ROCK signalling
Article Snippet: Paragraph title: RNA extraction and RT-qPCR ... RT-qPCR was performed as follows: 100 ng of cDNA was added to 10 µl of SYBR Premix Ex Taq II (Ti RNase H Plus; Takara Bio, Japan) and 200 nM of each primer, in a final volume of 20 µl; the PCR product was run on the Roche LightCycler 480 as follows: 95 °C for 2 min, then 40 cycles of 95 °C for 30 s, 60 °C for 1 min, followed by a melt curve analysis; and RT-qPCR was performed for each sample with each primer pair in triplicate.

Quantitation Assay:

Article Title: Redirecting abiraterone metabolism to fine tune prostate cancer anti-androgen therapy
Article Snippet: SYBR Premix Ex Taq II (Takara) was used for SRD5A1 , SRD5A2 and AR v7 detection. .. Accurate quantitation of each mRNA was achieved by normalizing the sample values to RPLPO and to vehicle-treated cells.

Spectrophotometry:

Article Title: Bacterial Communities Changes during Food Waste Spoilage
Article Snippet: Quantitative PCR was performed in a 25-μL reaction mixture containing 12.5 μL SYBR Premix Ex Taq II (Tli RNase H Plus, 2×; Takara), 0.48 μM each primer, and 2 μL template DNA. .. Standard plasmid DNA was prepared with a Plasmid Mini Kit (Omega), and its concentration was determined using a NanoDrop 2000 UV–vis spectrophotometer (Thermo Scientific, Wilmington DE, USA).

Concentration Assay:

Article Title: Hydrogen sulfide upregulated mRNA expressions of sodium bicarbonate cotransporter1, trefoil factor1 and trefoil factor2 in gastric mucosa in rats
Article Snippet: The purity and concentration of the extracted RNA was determined spectrophotometerically at 260 and 280 nm wavelength (BioPhotometer Plus; Eppendorf, Hamburg, Germany). .. All PCR amplifications were performed in duplicate reactions and in final volume of 20 µL containing 2 µL cDNA, 0.8 µL of specific primers, 10.0 µL of master SYBR Green Master Mix: SYBR Premix Ex Taq II (Tli RNase H Plus; Takara Bio Inc., Shiga, Japan), 6.4 µL ddH2 O using the following protocol: Pre-incubation at 95 ˚C for 2 min to activate DNA Taq polymerase and 40 two-step cycles with denaturation with denaturation at 95 ˚C for 20 sec, and annealing/ extension at 60 ˚C for 1 min.

Article Title: Bacterial Communities Changes during Food Waste Spoilage
Article Snippet: Quantitative PCR was performed in a 25-μL reaction mixture containing 12.5 μL SYBR Premix Ex Taq II (Tli RNase H Plus, 2×; Takara), 0.48 μM each primer, and 2 μL template DNA. .. Standard plasmid DNA was prepared with a Plasmid Mini Kit (Omega), and its concentration was determined using a NanoDrop 2000 UV–vis spectrophotometer (Thermo Scientific, Wilmington DE, USA).

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  • 90
    TaKaRa sybr premix ex taq ii
    Sybr Premix Ex Taq Ii, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 3775 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr premix ex taq ii/product/TaKaRa
    Average 90 stars, based on 3775 article reviews
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    sybr premix ex taq ii - by Bioz Stars, 2020-01
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