Tb green premix ex taq 2 tli rnaseh plus kit

Manufactured by Takara Bio

Sourced in China, Japan, United States, Germany

The TB Green® Premix Ex Taq™ II (Tli RNaseH Plus) kit is a reagent for real-time PCR analysis. It contains a DNA polymerase, buffer, dNTPs, and a fluorescent dye for detecting and quantifying target DNA sequences.

Automatically generated - may contain errors

Lab products found in correlation

Steponeplus real time pcr system, by Thermo Fisher Scientific (11 mentions) Trizol reagent, by Thermo Fisher Scientific (7 mentions) Primescript rt master mix, by Takara Bio (5 mentions) Cfx96 real time pcr detection system, by Bio-Rad (4 mentions) Primescript rt reagent kit, by Takara Bio (4 mentions) Rnaiso plus, by Takara Bio (4 mentions) Rneasy mini kit, by Qiagen (4 mentions) Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (3 mentions) Primescript rt master mix perfect real time kit, by Takara Bio (3 mentions) Primescript rt reagent kit with gdna eraser, by Takara Bio (3 mentions) Lightcycler 96, by Roche (3 mentions) Quantstudio 3 real time pcr system, by Thermo Fisher Scientific (3 mentions) Trizol reagent, by Takara Bio (2 mentions) Primescript rt master mix kit, by Takara Bio (2 mentions) Cfx connect real time pcr detection system, by Bio-Rad (2 mentions) Primescript rt reagent kit with gdna eraser perfect real time, by Takara Bio (2 mentions) Rox reference dye, by Bio-Rad (2 mentions) Ddh2o, by Bio-Rad (2 mentions) Tb green premix ex taq 2, by Takara Bio (2 mentions) Minibest universal rna extraction kit, by Takara Bio (2 mentions)

62 protocols using tb green premix ex taq 2 tli rnaseh plus kit

Quantifying Viral Load and Host Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

The viral RNA was extracted from supernatant, and cDNA was synthesized. Virus production was quantified by measuring the expression of the PEDV M gene. The qPCR was performed with a TB Green® Premix Ex Taq™ II Kit (Tli RNaseH Plus) (Takara Bio) in a 20 μL reaction mixture containing the primers for PEDV M gene (Table 2) (Wongthida et al. 2017 (link)). The qPCR was performed with initial denaturation at 95 °C for 30 s, followed by 40 cycles at 95 °C for 5 s, and 58 °C for 34 s. To calculate the viral copy number, a recombinant plasmid pMD19-T-PEDV-M containing a truncated M gene fragment was constructed, and tenfold serially diluted pMD19-T-PEDV-M was used to generate a standard curve.
To analyze the expression of several IFNs, IRF3, and IRF7, TRIzol™ reagent was added to each well to lyse and homogenize the treated cells. The total RNA was extracted, and the cDNA was synthesized. The qPCR was performed with the TB Green® Premix Ex Taq™ II Kit (Tli RNaseH Plus) (Takara Bio) in a 20 μL reaction mixture containing the specific primers (Table 1). The two-step qPCR was performed as described above. The cellular housekeeping gene β-actin (ACTB) was used to normalize the individual samples. The relative quantification of gene expression was evaluated with the 2^−ΔΔCt method (Livak and Schmittgen 2001 (link)).

Wang F., Wang M., Zhang L., Cheng M., Li M, & Zhu J. (2022). Generation and functional analysis of single chain variable fragments (scFvs) targeting the nucleocapsid protein of Porcine epidemic diarrhea virus. Applied Microbiology and Biotechnology, 106(3), 995-1009.

+ Open protocol

+ Expand

Quantitative Real-Time PCR Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

qRT-PCR was performed using a TB Green Premix Ex Taq II kit (Tli RNaseH Plus) (Takara) according to the manufacturer’s instructions for gene expression analysis. The template cDNA fragment was synthesized from the extracted total RNA and was diluted to 1/10 for use in the qRT-PCR system. All the primers are detailed in Supplementary Table S1. The procedure was performed with a LightCycler 480II machine (Roche, Switzerland) using a 45-cycle program (95°C for 5 s and 60°C for 30 s). The relative expression of genes was calculated based on the threshold cycle according to the _{Δ Δ} Ct method (Schmittgen and Livak, 2008 (link)). The result was normalized to the expression of the reference gene actin in mulberry (Chen et al., 2018 ) or L25 in tobacco (Liu et al., 2018 (link)). Expression was determined for at least three replicates per treatment.

Sun H., Zhao W., Liu H., Su C., Qian Y, & Jiao F. (2020). MaCDSP32 From Mulberry Enhances Resilience Post-drought by Regulating Antioxidant Activity and the Osmotic Content in Transgenic Tobacco. Frontiers in Plant Science, 11, 419.

+ Open protocol

+ Expand

Temporal Transcriptomic Analysis of Tartary Buckwheat Grains

Check if the same lab product or an alternative is used in the 5 most similar protocols

Grains from Xiaomiqiao and Jinqiaomai 2 after pollination (5, 10, 15 and 20 days) were sampled and immediately frozen in liquid nitrogen and stored at − 80 °C. Samples from each stage consisted of three biological replicates. Total RNA was isolated from grains using the Tiangen Plant RNA Purification Kit (Beijing, China) according to the manufacturer’s instructions. RNA samples with an A260/A230 ratio ≥ 2.0 and A260/A280 ratio ≥ 1.8 were used to synthesize cDNA using the TAKARA PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (Dalian, China).
Quantitative RT-PCR was performed using the CFX96TM Real-Time PCR Detection System (Bio-Rad, USA). The PCRs (20 μL) were conducted according to the instructions for the TB Green® Premix Ex Taq™ II Kit (Tli RNaseH Plus) (RR820b, TAKARA, Dalian, China). All PCRs were performed in two biological replicates. The quantitative PCR conditions were as follows: 95 °C for 30 s, followed by 40 cycles of 95 °C for 5 s, 60 °C for 34 s, and 72 °C for 20 s. Data were analysed by the 2^−(ΔΔCt) method to obtain relative mRNA expression data. Primers used for qRT-PCR amplification of the candidate genes for hull type and of actin in Tartary buckwheat were designed using Primer Premier 6.0 and are listed in Additional file 7: Table S6.

Shi T.X., Li R.Y., Zheng R., Chen Q.F., Li H.Y., Huang J., Zhu L.W, & Liang C.G. (2021). Mapping QTLs for 1000-grain weight and genes controlling hull type using SNP marker in Tartary buckwheat (Fagopyrum tataricum). BMC Genomics, 22, 142.

+ Open protocol

+ Expand

Citrus Gene Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted from different tissues (leaves, stem, root, flower, peel, and pulp) of WT citrus, treated leaves of WT citrus, and the calluses and leaves of transgenic plants with a FastPure Plant Total RNA Isolation Kit (Vazyme Biotech, China) and reverse-transcribed into cDNA using the PrimeScript™ RT Reagent Kit (+ gDNA Eraser; TaKaRa Bio, China). Gene expression was measured using this cDNA and a TB Green Premix Ex Taq™ II Kit (Tli RNaseH Plus; TaKaRa Bio, China) on a QuantStudio 5 Real-Time PCR Cycler (Applied Biosystems, NIST, USA), adopting annealing temperatures determined based on the gene-specific primers. The thermocycler program was set as follows: initial denaturation at 95°C for 30 s, followed by 40 cycles of amplification at 95°C for 5 s and 60°C for 34 s, and a final step of 95°C for 15 s, 60°C for 1 min, and 95°C for 15 s. The relative fold changes in the expression of the target genes were determined following the 2^−ΔΔCt method, using CsActb as the reference gene [39 (link)]. The CsCYP82L1- and CsCYP82L2-specific primers used in this assay are given in Supplementary Data Table S3. All assays were carried out using three independent biological replicates per sample.

Sun X., Hu C., Yi G, & Zhang X. (2024). Identification and characterization of two P450 enzymes from Citrus sinensis involved in TMTT and DMNT biosyntheses and Asian citrus psyllid defense. Horticulture Research, 11(4), uhae037.

+ Open protocol

+ Expand

RNA Extraction and qPCR Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA from leaves was extracted using the GeneJET Plant RNA Purification Mini Kit (MBI Fermentas, Canada). RNA samples were run on 1% agarose gels, and the quality was checked using a NanoDrop2000 spectrophotometer. The cDNA was synthesized using the RevertAid First-Strand cDNA Synthesis Kit (MBI Fermentas, Canada), and cDNA was used for fluorescence quantitative PCR analysis with a TB Green Premix Ex Taq II kit (Tli RNaseH Plus) (TaKaRa, Japan) and actin gene was used as the reference gene.

Lv J., Chen Y.Q., Ding A.M., Lei B., Yu J., Gao X.M., Dai C.B, & Sun Y.H. (2021). Control of axillary bud growth in tobacco through toxin gene expression system. Scientific Reports, 11, 17513.

+ Open protocol

+ Expand

Quantitative Validation of Carotenoid and Photosynthesis Genes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Based on the transcriptome results, seven differentially expressed genes related to carotenoid metabolism, fatty acid metabolism, photosynthesis, and cellular antioxidant capacity were selected for quantitative validation of gene expression using qRT-PCR. The internal reference gene was designed according to the large subunit of the ribulose 1,5-bisphosphate carboxylase/oxygenase gene (Rbcl).
RNA extraction was conducted using the E.Z.N.A.^® Plant RNA Kit (OMEGA, Germany), and the integrity of the 28S and 18S bands was checked by agarose gel electrophoresis to verify the RNA quality. The total RNA was reverse transcribed into cDNA, and the nucleic acid concentration was determined by Nanodrop. qPCR was performed using the TB Green Premix Ex Taq II Kit (Tli RNaseH Plus) (Takara, Dalian, China), and each sample was set up in triplicate. The experimental data were analyzed and plotted according to the RQ = 2⁻^△△Ct. qPCR genes and primers are shown in Table 2.

Sun J., Zan J, & Zang X. (2022). Research of Fluridone’s Effects on Growth and Pigment Accumulation of Haematococcus pluvialis Based on Transcriptome Sequencing. International Journal of Molecular Sciences, 23(6), 3122.

+ Open protocol

+ Expand

RNA Extraction and qRT-PCR Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted using TRIZOL reagent (Invitrogen). Subsequently, cDNA synthesis was performed in a reaction mixture consisting of M-MLV Reverse Transcriptase, RNase inhibitor, dNTPs, and Recombinant RNase inhibitor (all from Promega) at 42°C for 1 h. The cDNA samples were assayed in duplicate for qRT-PCR using the TB Green Premix Ex Taq II kit (Tli RNaseH Plus) (Takara Bio), according to the manufacturer’s instructions. All reactions were conducted in triplicate, and the expression levels were normalized to that of the 18s rRNA gene. The amplification process involved a reaction cycle comprising 95°C for 30 sec, followed by 95°C for 5 sec and 60°C for 30 sec. The fluorescence signal was detected at the end of the cycle using the LightCycler480 II instrument (Roche). mRNA expression levels were normalized to that of 18s rRNA using the 2^–ΔΔCT method [21 (link)]. The primer sequences used are: HIF-1α forward: 5ʹ- ACCACCTATGACCTGCTTGG-3ʹ and reverse: 5ʹ-CATATCCAGGCTGTGTCGAC-3ʹ; ZEB1 forward: 5ʹ-TTCACAGTGGAGAGAAGCCA-3ʹ and reverse: 5ʹ-GCCTGGTGATGCTGAAAGAG-3ʹ; 18s rRNA forward: 5ʹ-TAGTCGCCGTGCCTACCA-3ʹ and reverse: 5ʹ-TGCTGCCTTCCTTGGATGT-3ʹ.

Park S.J., Lee N, & Jeong C.H. (2024). ACY-241, a histone deacetylase 6 inhibitor, suppresses the epithelial–mesenchymal transition in lung cancer cells by downregulating hypoxia-inducible factor-1 alpha. The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology, 28(1), 83-91.

+ Open protocol

+ Expand

Quantitative Analysis of Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total cellular RNA was extracted using an EZ-press RNA Purification Kit (HiFunBio, Shandong, China) according to the manufacturer’s instructions. After determining RNA concentrations in samples using a fluorescence spectrometer, reverse transcription of 800 ng mRNA was performed using a First Strand cDNA Synthesis Kit (Qiagen, Germany). TB green Premix Ex Taq II kit (Tli RNase H Plus; Takara, Japan) to assess via RT-qPCR the expression of target genes, including COL22A1, IGF2BP2, MPO, and ACTB, with relative expression levels normalized to ACTB. The following primers were used: COL22A1 (forward: 5’-TCCGACTTCAATGCCATCGAC-3’; reverse: 5’- TACACGAACGCTAGGACAGAG-3’); IGF2BP2 (forward: 5’-AGTGGAATTGCATGGGAAAATCA-3’; reverse: 5’-CAACGGCGGTTTCTGTGTC-3’); MPO (forward: 5’-TGCTGCCCTTTGACAACCTG-3’; reverse: 5’-TGCTCCCGAAGTAAGAGGGT -3’); and ACTB (forward: 5’-CATGTACGTTGCTATCCAGGC-3’; reverse: 5’-CTCCTTAATGTCACGCACGAT-3’).

Liu H., Zeng Z, & Sun P. (2023). Prognosis and immunoinfiltration analysis of angiogene-related genes in grade 4 diffuse gliomas. Aging (Albany NY), 15(18), 9842-9857.

+ Open protocol

+ Expand

Quantitative PCR Analysis of Bacterial Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Culture and RNA extraction of bacteria were described above. Residual gDNA was digested and cDNA was synthesized using the HiScript II 1st Strand cDNA Synthesis Kit (+ gDNA wiper; Nanjing Vazyme Biotech Co., Ltd., China). Using the obtained cDNA as template, qPCR was performed with TB Green Premix Ex Taq II Kit (Tli RNaseH Plus) (Takara Biomedical Technology (Beijing) Co., Ltd., China) using primers of differentially expressed genes listed in Additional file 3. The 2^−ΔΔCt of each gene was calculated as the relative expression level with the 16S rRNA expression amount as the endogenous reference.

Zhang Q., Peng L., Han W., Chen H., Tang H., Chen X., Langford P.R., Huang Q., Zhou R, & Li L. (2023). The morphology and metabolic changes of Actinobacillus pleuropneumoniae during its growth as a biofilm. Veterinary Research, 54, 42.

+ Open protocol

+ Expand

Quantifying MSCs Pluripotency Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted using the TRIzol reagent (Life Technologies, USA) according to the manufacturer’s instructions. RNA purity and concentration were measured using a spectrophotometer (Epoch 2; BioTek, USA). cDNA was synthesized from RNA using a NICSROgene reverse transcription kit (Bionics, Korea). To identify the expression levels of MSCs pluripotency markers (OCT4, SOX2, and NANOG), real-time reverse transcription-polymerase chain reaction was performed using the TB Green Premix Ex Taq II Kit (Tli RnaseH Plus; TaKaRa, Japan) in a CFX-96 Real-Time PCR Detection System (Bio-Rad, USA). Thermocycler settings were 95°C for 30 sec, 40 cycles at 95°C for 5 sec, 60°C for 30 sec, and 95°C for 5 sec. Sequences for all used primers (Bionics) are listed in Table 1. The experiments were performed in triplicates. All results were standardized to glyceraldehyde 3-phosphate dehydrogenase expression. The relative level of mRNA expression was calculated using the 2 ^−ΔCt method.

Park M.K, & Song K.H. (2024). Isolation and characterization of feline endometrial mesenchymal stem cells. Journal of Veterinary Science, 25(2), e31.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!