Abi viia 7 dx system

Manufactured by Thermo Fisher Scientific

Sourced in United States

The ABI ViiA 7 DX system is a real-time PCR instrument designed for quantitative gene expression analysis and genotyping. The system features seven optical channels, a thermal block that can accommodate 96-well plates, and advanced data analysis software. It is intended for use in research and diagnostic laboratory settings.

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Lab products found in correlation

Primescript rt reagent kit, by Takara Bio (3 mentions) Rnaiso plus, by Takara Bio (2 mentions) Sybr premix ex taq 2, by Takara Bio (2 mentions) Nanodrop, by Thermo Fisher Scientific (2 mentions) Primescrip rt reagent kit with gdna eraser, by Takara Bio (1 mentions) Tb green premix ex taq 2, by Takara Bio (1 mentions) Rnaprep pure plant kit, by Tiangen Biotech (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Power sybr green master mix, by Thermo Fisher Scientific (1 mentions) Sybr premix ex taq, by Takara Bio (1 mentions) Minibest plant rna extraction kit, by Takara Bio (1 mentions) Takara minibest plant rna extraction kit, by Takara Bio (1 mentions) Nanodrop system, by Implen (1 mentions) Primescrip rt reagent kit with gdna eraser perfect real time, by Takara Bio (1 mentions) Reverse transcription reagent kit, by Takara Bio (1 mentions) Sybr green qpcr master mix, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

ABI ViiA 7 Dx ViiATM 7 Dx system ABI ViiA 7 system ViiA 7 Dx ABI ViiA 7

8 protocols using abi viia 7 dx system

Quantification of Gene Expression in Kidney Tissues

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RNAiso Plus (TaKaRa Biotech, Dalian, China) was used to extract total RNA from frozen kidney tissues and HK-2 cells according to the instructions provided by the manufacturer. Subsequently, the PrimeScript™ RT Reagent Kit (TaKaRa Biotech) was used for reverse transcription into cDNA. In all PCR experiments, the expression of glyceraldehyde-3-phosphate dehydrogenase was used as the internal reference. The qRT-PCR analysis was performed using the ABI ViiA7DX System (Applied Biosystems, Foster City, CA, USA). The following qRT-PCR primers for the specific target genes (listed below) were designed and synthesized by TaKaRa Biotech:
R-Dot1l: 5‘-TGACCGCACCATACTTGAAAACTA-3ʹ(F)
5‘-GGCTCTTCTTGGACTTGCGAT-3ʹ(R);
R -β-actin: 5‘-TGCTATGTTGCCCTAGACTTCG-3ʹ(F)
5‘-GTTGGCATAGAGGTCTTTACGG-3ʹ(R);
H-α-SMA: 5‘-CCTATCCCCGGGACTAAGAC-3ʹ(F), 5‘-CCATCACCCCCTGATGTCTG-3ʹ(R);
H-Vimentin: 5‘-GGAGGAGATGCTTCAGAGAGAG-3ʹ(F), 5‘-GGATTTCCTCTTCGTGGAGTTTC-3ʹ(R);
H-Fibronectin: 5‘-GCCGAATGTAGGACAAGAAGC-3ʹ(F),
5‘- AAGCACGAGTCATCCGTAGGT-3ʹ(R);
H-β-actin: 5‘-CACCCAGCACAATGAAGATCAAGAT-3ʹ(F), 5‘-CCAGTTTTTAAATCCTGAGTCAAGC-3ʹ(R);
Routine qRT-PCR was performed as follows: 94°C for 3 mins, followed by 30 cycles (25 cycles for β-actin) at 94°C for 30 s, 55°C for 30 s, and 72°C for 1 min.

Yang C., Chen Z., Yu H, & Liu X. (2019). Inhibition of Disruptor of Telomeric Silencing 1-Like Alleviated Renal Ischemia and Reperfusion Injury-Induced Fibrosis by Blocking PI3K/AKT-Mediated Oxidative Stress. Drug Design, Development and Therapy, 13, 4375-4387.

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Quantitative RNA Expression Analysis

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Total RNA was extracted with the RNAprep Pure Plant Kit (Tiangen Biotech Co., Ltd., Beijing, China). RNA integrity was evaluated using agarose gel electrophoresis and a NanoDrop spectrophotometer (Thermo Scientific, Los Angeles, CA, USA). A total of 1 μg high-quality RNA was used as the input material for cDNA synthesis with the PrimeScrip RT Reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Inc., Dalian, China). Real-time quantitative PCR (qRT-PCR) was implemented on the ABI ViiA 7 DX system (Applied Biosystems) using TB Green Premix Ex Taq II (TaKaRa) with the ubiquitin (UBQ) gene as a reference gene to normalize expression data. The specific primer used for qRT-PCR was designed using Primer Premier 5 software and the sequence is listed in Table S1. Each PCR reaction contained 5.0 µL TB Green mix, 0.8 µL primers, and 1.0 µL diluted cDNA in a final volume of 10 µL. The amplification conditions were as follows: 30 s of denaturation at 95 °C, followed by 40 cycles at 95 °C for 5 s and 60 °C for 30 s, then 95 °C for 15 s, 60 °C for 1 min, 95 °C for 15 s. Each experiment was repeated at least triplicate and each gene was calculated with the 2^−ΔΔCT method for the relative expression.

Luo K., Li J., Lu M., An H, & Wu X. (2023). Genome-Wide Identification and Expression Analysis of Rosa roxburghii Autophagy-Related Genes in Response to Top-Rot Disease. Biomolecules, 13(3), 556.

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Quantitative Transcriptome Analysis

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Total RNAs were extracted by TRIzol reagent (Invitrogen) and then reverse-transcribed to complementary DNA using the PrimeScript RT reagent kit (TaKaRa). qRT-PCR was performed using Power SYBR Green Master mix (Applied Biosystems) on an ABI Viia7 DX System (Applied Biosystems). Primers are listed in Supplementary Table S7.

Feng Y.L., Xiang J.F., Liu S.C., Guo T., Yan G.F., Feng Y., Kong N., Li H.D., Huang Y., Lin H., Cai X.J, & Xie A.Y. (2017). H2AX facilitates classical non-homologous end joining at the expense of limited nucleotide loss at repair junctions. Nucleic Acids Research, 45(18), 10614-10633.

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RNA Isolation and RT-qPCR Analysis in NRK-52E Cells

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Total RNA was isolated from NRK-52E cells using RNAiso Plus (Takara Bio, Inc.) according to the manufacturer's instructions and subjected to reverse transcription into cDNA using the PrimeScript™ RT Reagent kit (Takara Bio, Inc.) Reverse transcription was performed at 37°C for 15 min and then 95°C for 5 sec. RT-qPCR analysis was performed using SYBR^® Premix Ex Taq™ (Takara Bio, Inc.) and the ABI ViiA7DX System (Applied Biosystems; Thermo Fisher Scientific, Inc.). β-actin expression levels were used as an internal reference for all PCR experiments. RT-qPCR primers designed for specific target genes were synthesized by Takara Bio, Inc. (Table SI). PCR reactions were performed using the following cycling conditions: 95°C for 30 sec, followed by 40 cycles at 95°C for 5 sec, 60°C for 30 sec and 72°C for 20 sec. The relative mRNA levels for each sample were calculated by the 2^−ΔΔcq method (23 (link)).

Yin X., Ma F., Fan X., Zhao Q., Liu X, & Yang Y. (2020). Knockdown of AMPKα2 impairs epithelial-mesenchymal transition in rat renal tubular epithelial cells by downregulating ETS1 and RPS6KA1. Molecular Medicine Reports, 22(6), 4619-4628.

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Plant RNA Extraction and qRT-PCR Analysis

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Total RNA was extracted using a TaKaRa MiniBEST Plant RNA Extraction Kit (TaKaRa, Dalian, China). cDNA was synthesized after removing the genomic DNA with the PrimeScrip RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Dalian, China). qRT-PCR was performed on an ABI ViiA 7 DX system (Applied Biosystems) using SYBR Premix Ex Taq II (TaKaRa, Dalian, China). Transcript levels were normalized to RrUBQ (ubiquitin, internal control gene, ID: evm.model.Contig289.99) (Lu M. et al., 2020 (link)), and data analysis was performed using the 2^−ΔΔCT method (Livak and Schmittgen, 2001 (link)). Values for mean expression and standard deviation (SD) were calculated from the results of three independent experiments. The primer sequences used for qRT-PCR are listed in Supplemental Table 5.

Yan Y., Liu Y., Lu M., Lu C., Ludlow R.A., Yang M., Huang W., Liu Z, & An H. (2023). Gene expression profiling in Rosa roxburghii fruit and overexpressing RrGGP2 in tobacco and tomato indicates the key control point of AsA biosynthesis. Frontiers in Plant Science, 13, 1096493.

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Validating Dietary Fiber DEGs via qRT-PCR

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Candidate differentially expressed genes (DEGs) involved in dietary fiber metabolism were selected for validation by real time quantitative PCR (qRT-PCR). Total RNA was extracted through a TaKaRa MiniBEST Plant RNA Extraction Kit (TaKaRa, Inc., Dalian, China). RNA quality was evaluated by agarose gel electrophoresis and the NanoDrop system (Implen, Los Angeles, CA, USA). cDNA was synthesized with the PrimeScrip RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Inc., Dalian, China). The primer sequences used for qRT-PCR are listed in Table S1. qRT-PCR was performed on an ABI ViiA 7 DX system (Applied Biosystems) using SYBR Premix Ex Taq II (TaKaRa) with the ubiquitin gene as an endogenous control. Data analysis was performed using the 2 -ΔΔCT method. Values for mean expression and standard deviation (SD) were calculated from the results of three independent experiments.

Zhang X., Lu M., Ludlow R.A., Ma W., & An H. (2021). Transcriptome analysis reveals candidate genes for dietary fiber metabolism in Rosa roxburghii fruit grown under different light intensities. Horticulture Environment and Biotechnology, 62(5), 751-764.

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qPCR Analysis of P. aeruginosa RNA

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The RNA of P. aeruginosa strains was extracted for qPCR analysis according to the RNA reagent instructions (Promega, Madison, WI, USA): (I) 1 μg of the extracted RNA was reverse transcribed into cDNA with a reverse transcription reagent kit (TaKaRa, Dalian, Liaoning, China); (II) following the instructions of the SYBR Green qPCR Master Mix (Thermo Fisher Scientific, Waltham, MA, USA), relative real-time PCR was performed with the ABI ViiA^TM 7Dx system (Applied Biosystems, Foster, CA, USA) to detect cDNA; (III) the housekeeping gene ribosomal protein rpoD was used as an internal control for normalization. Relative changes in gene expression levels were calculated with the 2^−ΔΔCt method. Primer sequences are shown in Table 1.

Xiao Q., Luo Y., Shi W., Lu Y., Xiong R., Wu X., Huang H., Zhao C., Zeng J, & Chen C. (2022). The effects of LL-37 on virulence factors related to the quorum sensing system of Pseudomonas aeruginosa. Annals of Translational Medicine, 10(6), 284.

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Profiling Ubiquitination Pathway Genes

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The Human Ubiquitination Pathway RT² Profiler PCR Array (Cat. no. 330231, Qiagen, Hilden, German) was used to profile the expression of 84 key genes involved in the regulated degradation of cellular proteins by the ubiquitin-proteasome. Total RNA was extracted from SW620 and SW48 with TRIzol. Single stranded cDNA was synthesized from 2 μg of total RNA by using the SuperArray reaction ready first strand cDNA synthesis kit. The cDNAs were mixed with SuperArray RT² Real time SYBR Green/ROX PCR master mix and real-time PCR performed in accordance with the manufacturer’s instructions. Thermal cycling and fluorescence detection were performed using an ABI ViiA^TM 7 Dx System (Applied Biosystems, Foster City, CA, USA), and expression of these 84 E3 ubiquiting ligases gene transcripts was compared between SW620 and SW48.

Liu S., Fei W., Shi Q., Li Q., Kuang Y., Wang C., He C, & Hu X. (2017). CHAC2, downregulated in gastric and colorectal cancers, acted as a tumor suppressor inducing apoptosis and autophagy through unfolded protein response. Cell Death & Disease, 8(8), e3009-.

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