Abi quantstudio 7 detection system

Manufactured by Thermo Fisher Scientific

Sourced in United States

The ABI Quantstudio 7 detection system is a real-time PCR instrument designed for sensitive and precise nucleic acid quantification. It features a 96-well format and supports multiple detection chemistries, enabling a wide range of gene expression and genetic analysis applications.

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Lab products found in correlation

Powerup sybr green master mix, by Thermo Fisher Scientific (3 mentions) Nanodrop spectrophotometer, by Thermo Fisher Scientific (2 mentions) Elisa max standard set mouse tnf α, by BioLegend (1 mentions) Mouse il 6 elisa max standard set, by BioLegend (1 mentions) Power up sybr green kit, by Thermo Fisher Scientific (1 mentions) Primescript rt reagent kit with gdna eraser, by Takara Bio (1 mentions) M5 easyspin plus kit, by Mei5 Biotechnology (1 mentions) Hipure total rna plus mini kit, by Magen Biotechnology Co (1 mentions) Hipure bacterial dna kit, by Magen Biotechnology Co (1 mentions) Primescript rt reagent kit, by Takara Bio (1 mentions) 6 well plate, by Corning (1 mentions)

4 protocols using abi quantstudio 7 detection system

Cytokine Production Regulation in MH-S Cells

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MH‐S cells were plated in 12‐well plates at a density of 7.5 × 10⁵ cells per well. Cells were treated with 50 µg mL⁻¹ of SLAP‐S25, GNGs, and 10 µm dexamethasone for 30 min before the final concentration of LPS (1 µg mL⁻¹) was added to cell cultures. After 4 h of incubation, the supernatants were obtained for TNF‐α and IL‐6 production analysis using ELISA MAX Standard Set Mouse TNF‐α and ELISA MAX Standard Set Mouse IL‐6 (Biolegend, US).
The expression of TNF‐α and IL‐6 relative to β‐actin was detected by qRT‐PCR tests with the PowerUp SYBR Green Kit (Applied Biosystems). Thermal cycling was performed using a two‐step PCR amplification standard procedure at 95 °C for 30 s and 40 cycles of 60 °C for 30 s and 72 °C for 30 s. The qRT‐PCR test was performed using the ABI Quantstudio 7 detection system (Applied Biosystems). The fold changes of gene expression were determined using the 2^−ΔΔCt method.

Li X., Qu S., Song X., Wu C., Shen J, & Zhu K. (2023). In Situ Neutralization and Detoxification of LPS to Attenuate Hyperinflammation. Advanced Science, 10(26), 2302950.

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Bacterial RNA Expression Analysis

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Bacterial total RNA was extracted and examined using M5 EASYspin Plus kit (Mei5bio, Beijing, China) and Nanodrop spectrophotometer (Thermo Scientific, MA, United States), respectively. Reverse transcription was performed using a PrimeScript RT reagent Kit with gDNA Eraser (Takara, Beijing, China) with the manufacturer’s protocol. The messenger RNA levels relative to those of the control genes 16S were determined by real-time PCR tests with PowerUp SYBR Green Master Mix (Applied Biosystems, Thermo Fisher Scientific, Carlsbad, CA, United States). RT-PCR tests were performed using the ABI Quantstudio 7 detection system (Applied Biosystems, Thermo Fisher Scientific, Carlsbad, CA, United States). The fold changes in gene expression were determined using the 2^–ΔΔCt method. Primers used in this study were listed in Supplementary Table 3.

Gong X., Zhao Q., Wu Y., Zhou H., Ding S, & Zhu K. (2022). Mucoid Acinetobacter baumannii enhances anti-phagocytosis through reducing C3b deposition. Frontiers in Medicine, 9, 879361.

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Antioxidant Response in S. aureus Infection

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PCR analysis was performed to detect the antioxidants of S. aureus. Total DNA was extracted using the HiPure Bacterial DNA Kit (Magen). Primer sequences are listed in Table S5. RT-qPCR analysis was performed according to previously described research [23] . Briefly, RAW264.7 cells were seeded in an antibiotic-free medium at a concentration of 2 × 10⁶ cells per well in 6-well plates. Cells were treated with 8 μg mL^-¹ enrofloxacin for 12 h. Alternatively, macrophages were infected with S. aureus (MOI = 5) for 1 h, followed by the treatment of 8 μg mL^-¹ enrofloxacin for 12 h. Total RNA was extracted using the HiPure Total RNA Plus Mini Kit (Magen), and 1 μg of extracted RNA was studied using the PrimeScript RT Reagent Kit (Takara) according to the manufacturer's protocol. The messenger ribonucleic acid (mRNA) levels of mpo, nox1, nox2, nox3, sod1, sod2, trx1, trx2, cat, gpx1, il1b, il6, il10, ifnb1, tnfa, tgfb, mcp1, ifngr1, il4r, nos2, arg1, nrf2 herpud, atf4, xbp-1 s, chop and trib3 relative to that of the control gene actb were determined by real-time PCR tests with PowerUp SYBR Green Master Mix (Applied Biosystems). RT-qPCR was performed using an ABI QuantStudio 7 detection system (Applied Biosystems) for cycle threshold (Ct) values. The fold changes in gene expression were determined using the 2^-^ΔΔCt method. Primer sequences for RT-qPCR are listed in Table S6.

Wu Y., Gong X., Shen J, & Zhu K. (2023). Postantibiotic leukocyte enhancement-mediated reduction of intracellular bacteria by macrophages. Journal of Advanced Research, 58, 117-128.

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RT-qPCR Analysis of NAD+ Metabolic Genes

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RT-qPCR analyses were performed according to previously described research [60 (link)]. Briefly, MH-S and A549 cells were seeded in antibiotic-free medium at a concentration of 2 × 10⁶ cells per well in 6-well plates (Corning Incorporated Co., US). After that, the cells were infected with S. chromogenes SC10, SC1, SC2, SC5, SC14, or SC24; Av. paragallinarum X1-1S-1; S. aureus 12-1N-1; or S. warneri X1-2S-2 at an MOI of 1 or treated with 32 μg/mL vancomycin at 37°C for 6 h. Total RNA was extracted using a HiPure Total RNA Plus Mini Kit (Magen Biotechnology Co., Guangzhou, China), and 1 μg of extracted RNA was reverse transcribed with a PrimeScript RT Reagent Kit (TaKaRa Bio incorporated, Japan) following the manufacturer’s protocol. The messenger RNA levels of qprt, naprt, nadsyn1, nampt, nmrk1 and nmnat1 relative to those of the control genes gapdh or actb in MH-S or A549 cells were determined by real-time PCR tests with PowerUp SYBR Green Master Mix (Applied Biosystems, Thermo Fisher Scientific, US). RT-PCR tests were performed using the ABI Quantstudio 7 detection system (Applied Biosystems, Thermo Fisher Scientific, US). The fold changes in gene expression were determined using the 2^−ΔΔCt method. The primer sequences are listed in S6 Table.

Wu Y., Wang Y., Yang H., Li Q., Gong X., Zhang G, & Zhu K. (2021). Resident bacteria contribute to opportunistic infections of the respiratory tract. PLoS Pathogens, 17(3), e1009436.

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