Tb green premix ex taq kit tli rnase h plus

Manufactured by Takara Bio

Sourced in United States

The TB Green Premix Ex Taq kit (Tli RNase H Plus) is a reagent kit used in real-time PCR applications. The kit contains a premixed solution that includes the necessary components for efficient DNA amplification and detection using the TB Green dye.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (2 mentions) Ribospin plant kit, by GeneAll (1 mentions) Primescript rt reagent kit with gdna eraser, by Takara Bio (1 mentions) Dnase 1, by New England Biolabs (1 mentions) Primescript rt reagent kit, by Takara Bio (1 mentions) Primescript rt master mix kit, by Takara Bio (1 mentions) Lightcycler 96 instrument, by Roche (1 mentions) Primescript rt reagent kit with gdna eraser perfect real time, by Takara Bio (1 mentions) Stratagene mx3005p thermocycler, by Agilent Technologies (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Primescript rt reagent kit with the gdna eraser, by Takara Bio (1 mentions) Cfx connect real time pcr detection system, by Bio-Rad (1 mentions) Minibest plant rna extraction kit, by Takara Bio (1 mentions)

Close spelling variants of this product

TB Green Premix Ex Taq II (Tli RNase H Plus) kits TB Green Premix Taq Premix Ex Taq TB Green Premix TB Green kit Ex Taq Premix Taq TB Green RNase H

6 protocols using tb green premix ex taq kit tli rnase h plus

Reverse Transcription PCR Protocol

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To perform mRNA reverse transcription PCR (RT-PCR), 10 μg of total RNA isolated by TRIzol reagent (Thermo Fisher Scientific) was firstly treated with deoxyribonuclease I (DNase I) [New England Biolabs (NEB)]. After reextraction, 1 μg of total RNA was converted to cDNA with the PrimeScript RT Reagent Kit (Takara). Semiquantitative RT-PCR was performed with Taq enzyme, and quantitative RT-PCR (qRT-PCR) was performed with a TB Green Premix Ex Taq kit (Tli RNase H Plus) (Takara). Elongation factor 1-α (EF-1α) or ACTIN2 was used as internal control, and primers used were listed in table S4.
To perform sRNA qRT-PCR analysis, 1 μg of total RNA isolated by TRIzol reagent was treated with DNase I (NEB). cDNA was synthesized using Moloney murine leukemia virus reverse transcriptase (NEB) primed by miRNA/U6-specific primers. qRT-PCR was performed with a TB Green Premix Ex Taq kit (Tli RNase H Plus) (Takara). U6 was used as internal control, and primers used were listed in table S4.

Li Q., Liu N., Liu Q., Zheng X., Lu L., Gao W., Liu Y., Liu Y., Zhang S., Wang Q., Pan J., Chen C., Mi Y., Yang M., Cheng X., Ren G., Yuan Y.W, & Zhang X. (2021). DEAD-box helicases modulate dicing body formation in Arabidopsis. Science Advances, 7(18), eabc6266.

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Quantifying Plant Secondary Metabolites

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Total RNA was extracted with GeneAll Ribospin™ Plant Kit (Cat. 307-150; Gene all, Seoul, Korea). The extracted RNA was quantified using a UV/Vis Nanodrop spectrophotometer (MicroDigital, Nabi, Seongnam, Korea). RNA purity was measured using the absorbance ratio of OD 260/280 and OD 260/230. The cDNA library was synthesized using the PrimeScript™ RT reagent Kit with gDNA Eraser (RR047A; TaKaRa, Tokyo, Japan) according to the manufacturer’s protocol with 1 μg of RNA. The quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was conducted using the TB Green® Premix Ex Taq™∥ Kit (Tli RNaseH Plus) (RR820A; TaKaRa, Tokyo, Japan). The PCR primer sequences are listed in Supplementary Table 2. qRT-PCR was performed using the 12 genotypes of which the secondary metabolite contents varied significantly.

Kim B.C., Lim I, & Ha J. (2023). Metabolic profiling and expression analysis of key genetic factors in the biosynthetic pathways of antioxidant metabolites in mungbean sprouts. Frontiers in Plant Science, 14, 1207940.

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Confirming RNA-Seq Results by qRT-PCR

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Confirmation of RNA-Seq results by qRT-PCR was based on ten genes selected randomly from the comparison of CS vs. NT (Zjn_sc00026.1.g00600.1.am.mkhc, Zjn_sc00014.1.g01180.1.sm.mkhc, Zjn_sc00016.1.g04050.1.sm.mkhc, Zjn_sc00007.1.g00600.1.am.mkhc, Zjn_sc00086.1.g01640.1.am.mk) and CE vs. NT (Zjn_sc00133.1.g00260.1.am.mkhc, Zjn_sc00003.1.g01070.1.sm.mkhc, Zjn_sc00097.1.g01050.1.am.mk, Zjn_sc00017.1.g04240.1.sm.mkhc, Zjn_sc00016.1.g04940.1.am.mkhc). The cDNA preparations synthesized from the RNA samples were also used for qRT-PCR analysis based on ten randomly selected genes. Three technical repeats were processed for each of the three biological repeats originally performed per treatment. The primers were designed and synthesized by RuiBiotech Co., Ltd. (http://www.ruibiotech.com/, accessed on 2 February 2022) (Table 2). The instructions for the TB Green Premix Ex Taq kit (Tli RNaseH Plus) were obtained from Takara Bio Inc. (https://www.takarabiomed.com.cn/, accessed on 2 February 2022), and the qRT-PCR was performed using a Bio Rad CFX Connect Real-Time PCR Detection System (Bio-Rad Laboratories Co., Ltd., California, USA) (https://www.bio-rad.com/, accessed on 2 February 2022) with the following parameter settings: 95 °C for 30 s, followed by 40 cycles of 95 °C for 5 s and annealing/extension at 60 °C for 30 s. Single-product amplification was confirmed with a melt curve.

Zhang J., Zhang Z., Liu W., Li L., Han L., Xu L, & Zhao Y. (2022). Transcriptome Analysis Revealed a Positive Role of Ethephon on Chlorophyll Metabolism of Zoysia japonica under Cold Stress. Plants, 11(3), 442.

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Osteogenesis Induction RNA Extraction

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After 7 days of osteogenesis induction, RNAiso Plus (Takaro, Japan) was used to extract total RNA of different groups according to the manufacturer’s instructions. Applying PrimeScript™ RT Master Mix kit (Perfect Real Time) (Takara, Japan) to reverse transcription. qPCR was performed via the TB Green Premix Ex Taq™ kit (Tli RNaseH Plus) (Takara) in LightCycler@ 96 Instrument (Roche). β-tubulin was standardized as the internal reference. Primer sequences used in this study were listed in Table 1.Each sample was assayed in triplicate. Data analysis adopted 2^−ΔΔCT calculation method.

Lao A., Chen Y., Sun Y., Wang T., Lin K., Liu J, & Wu J. (2022). Transcriptomic analysis provides a new insight: Oleuropein reverses high glucose-induced osteogenic inhibition in bone marrow mesenchymal stem cells via Wnt10b activation. Frontiers in Bioengineering and Biotechnology, 10, 990507.

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Quantitative expression analysis of T. annulata

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Total RNA was extracted from T. annulata at three life-cycle stages (schizont, sporozoite and merozoite) using an RNeasy Mini Kit (QIAGEN, Hilden, Germany) according to the user manual. The complementary DNA (cDNA) templates were then synthesized using the PrimeScript™ RT Reagent Kit with gDNA Eraser (Perfect Real Time; TaKaRa, Bio Inc., Kusatsu, Shi, Japan) following evaluation of the concentration and quality of RNA. The specific primers for the target genes were designed according to the sequences derived from the NCBI database. The levels of the β-actin transcript in T. annulata and bovine cells were used for normalization. All primer sequences used in the present study are shown in Additional file 1: Table S1. Quantitative real-time PCR assays were performed in the Stratagene MX3005P thermocycler (Agilent Technologies, Santa Clara, CA, USA) and using the TB Green® Premix Ex Taq™ kit (Tli RNaseH Plus; TaKaRa Bio Inc.) according to the user manual. The relative transcript levels of the target genes were calculated using the comparative cycle threshold (2^−ΔΔCT) method.

Li Z., Liu J., Zhao S., Ma Q., Guo Z., Liu A., Li Y., Guan G., Luo J, & Yin H. (2022). Theileria annulata SVSP455 interacts with host HSP60. Parasites & Vectors, 15, 308.

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Quantification of Z. japonica Gene Expression

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The TaKaRa MiniBEST Plant RNA Extraction Kit (TaKaRa, Japan-b) (Beijing, China) was used to extract the total RNA of Z. japonica through grinding, splitting, cleaning, digestion, elution, and other steps, while the PrimeScriptTMRT reagent Kit with the gDNA Eraser (Takara, Japan-a) (Beijing, China) was used to synthesize the cDNA and remove the genomic DNA for a subsequent determination of gene expression. The primers were designed and synthesized by RuiBiotech Co., Ltd., Beijing, China (http://www.ruibiotech.com/) (accessed on 27 December 2023). The instructions for the TB Green Premix Ex Taq kit (Tli RNaseH Plus) were obtained from Takara Biomedical Technology (Beijing) Co., Ltd., Beijing, China (https://www.takarabiomed.com.cn/) (accessed on 27 December 2023), and qRT-PCR was performed using a Bio Rad CFX Connect Real-Time PCR Detection System (Bio-Rad Laboratories Co., Ltd., CA, USA) (https://www.bio-rad.com/) with the following parameter settings: 95 °C for 30 s, followed by 40 cycles of 95 °C for 5 s and annealing/extension at 60 °C for 30 s. Single-product amplification was confirmed with a melt curve. Relative gene expression was based on the 2^−ΔΔCT method [89 (link)], which is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments.

Zhang J., Li L., Zhang Z., Han L, & Xu L. (2024). The Effect of Ethephon on Ethylene and Chlorophyll in Zoysia japonica Leaves. International Journal of Molecular Sciences, 25(3), 1663.

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