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7500 fast dx

Manufactured by Thermo Fisher Scientific
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The 7500 Fast (Dx) is a real-time PCR system designed for diagnostic applications. It provides fast, accurate, and reliable detection of target nucleic acid sequences. The system is capable of performing thermal cycling and real-time fluorescence detection to enable quantitative analysis of DNA and RNA samples.

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Lab products found in correlation

Quantstudio 5, by Thermo Fisher Scientific (2 mentions) Taqpath 1 step rt qpcr master mix, by Thermo Fisher Scientific (2 mentions) Power sybr green pcr master mix, by Thermo Fisher Scientific (2 mentions) Microamp optical adhesive film, by Thermo Fisher Scientific (1 mentions) Microamp optical 96 well reaction plate, by Thermo Fisher Scientific (1 mentions) Taqman environmental master mix 2, by Thermo Fisher Scientific (1 mentions) Quantstudio 3, by Thermo Fisher Scientific (1 mentions) Tripure isolation reagent, by Merck Group (1 mentions) One step tb green primescript rt pcr kit 2, by Takara Bio (1 mentions) Amfirivert cdna synthesis platinum master mix, by GenDEPOT (1 mentions) Dnazol reagent, by Thermo Fisher Scientific (1 mentions) Human mitochondrial dna mtdna monitoring primer set, by Takara Bio (1 mentions)

7 protocols using 7500 fast dx

1

Standardized Multiplex qPCR Protocol

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All participants received standardized protocols, including detailed instructions for completing the study. Laboratories were supplied with sterile PCR-grade OmniPur water (VWR, Radnor, PA), 1.7 ml Axygen™ MaxyClear microcentrifuge tubes (Thermo Fisher Scientific, Grand Island, NY), SRM 2917 (NIST, Gaithersburg, MD), internal amplification control (IAC) for multiplex HF183/BacR287, HumM2, and CowM2 qPCR assays (102 copies/2 μL), a 1.5 mL aliquot of 2 mg/ml bovine serum albumin fraction V stock solution (Thermo Fisher Scientific), primer/hydrolysis probe stock solutions for each qPCR assay (Table 1), MicroAmp™ optical 96-well reaction plates (Thermo Fisher Scientific), MicroAmp™ optical adhesive film (Thermo Fisher Scientific), and TaqMan™ Environmental Master Mix 2.0 (Thermo Fisher Scientific). Participants were required to use a StepOnePlus™, 7500, 7500 Fast Dx, 7500 Fast™, QuantStudio™ 3, QuantStudio™ 5, or QuantStudio™ 7 Pro Dx real-time PCR system (Thermo Fisher Scientific). Using the required supplies, participants were instructed to (i) generate six calibration curves for each qPCR assay on separate instrument runs and (ii) submit data to the EPA (Cincinnati, OH). Participants were instructed to perform all experiments within 30 days.
Sivaganesan M., Willis J.R., Karim M., Babatola A., Catoe D., Boehm A.B., Wilder M., Green H., Lobos A., Harwood V.J., Hertel S., Klepikow R., Howard M.F., Laksanalamai P., Roundtree A., Mattioli M., Eytcheson S., Molina M., Lane M., Rediske R., Ronan A., D’Souza N., Rose J.B., Shrestha A., Hoar C., Silverman A.I., Faulkner W., Wickman K., Kralj J.G., Servetas S.L., Hunter M.E., Jackson S.A, & Shanks O.C. (2022). Interlaboratory performance and quantitative PCR data acceptance metrics for NIST SRM® 2917. Water research, 225, 119162.
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2

Quantifying Gene Expression via RT-qPCR

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Total RNA was purified using TriPure isolation reagent (Sigma-Aldrich) according to the manufacturer’s instructions. Total RNA was subjected to RT-qPCR using One Step TB Green PrimeScript RT-PCR Kit II (Takara) and real-time PCR system 7500 Fast Dx (ThermoFisher Scientific). The primers used for RT-qPCR are presented in Table 2.
Sagou K., Sato Y., Okuno Y., Watanabe T., Inagaki T., Motooka Y., Toyokuni S., Murata T., Kiyoi H, & Kimura H. (2024). Epstein-Barr virus lytic gene BNRF1 promotes B-cell lymphomagenesis via IFI27 upregulation. PLOS Pathogens, 20(2), e1011954.
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3

RT-qPCR Assay for SARS-CoV-2 Quantification

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We conducted RT-qPCR analysis using 20 μL reactions consisting of 5 μL template, 7 μL nuclease-free water, 5 μL TaqPath 1-Step RT-qPCR Master Mix (4×), and 1.5 μL of each N1 and N2 primer/probe mix from the 2019-nCoV CDC EUA kit (42 ) at a final concentration of 1.125 μM. We performed analysis using the Applied Biosystems 7500 Fast (Dx) real-time PCR system with a cycling condition of 2 min at 95°C followed by 45 cycles of 3 s at 95°C and 30 s at 55°C. SARS-CoV-2 RNA quantification was conducted using a 5-point standard curve generated from serial dilutions of the CDC EUA 2019-nCoV positive control, ranging from 20 copies/rxn to 1 × 105 copies/rxn. The required standard curve criteria were an efficiency between 80 and 120% and R2 greater than 0.98. We monitored the stability, method performance, and quantitative consistency by recording the CT of the 1 × 105 copies/rxn standard well longitudinally at a predetermined threshold.
Gupta P., Liao S., Ezekiel M., Novak N., Rossi A., LaCross N., Oakeson K, & Rohrwasser A. (2023). Wastewater Genomic Surveillance Captures Early Detection of Omicron in Utah. Microbiology Spectrum, 11(3), e00391-23.
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4

RT-qPCR Assay for SARS-CoV-2 Quantification

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We conducted RT-qPCR analysis using 20 μL reactions consisting of 5 μL template, 7 μL nuclease-free water, 5 μL TaqPath 1-Step RT-qPCR Master Mix (4×), and 1.5 μL of each N1 and N2 primer/probe mix from the 2019-nCoV CDC EUA kit (42 ) at a final concentration of 1.125 μM. We performed analysis using the Applied Biosystems 7500 Fast (Dx) real-time PCR system with a cycling condition of 2 min at 95°C followed by 45 cycles of 3 s at 95°C and 30 s at 55°C. SARS-CoV-2 RNA quantification was conducted using a 5-point standard curve generated from serial dilutions of the CDC EUA 2019-nCoV positive control, ranging from 20 copies/rxn to 1 × 105 copies/rxn. The required standard curve criteria were an efficiency between 80 and 120% and R2 greater than 0.98. We monitored the stability, method performance, and quantitative consistency by recording the CT of the 1 × 105 copies/rxn standard well longitudinally at a predetermined threshold.
Gupta P., Liao S., Ezekiel M., Novak N., Rossi A., LaCross N., Oakeson K, & Rohrwasser A. (2023). Wastewater Genomic Surveillance Captures Early Detection of Omicron in Utah. Microbiology Spectrum, 11(3), e00391-23.
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5

Comprehensive RNA Extraction and qRT-PCR Analysis

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Total RNA was extracted using a commercial RNA extraction kit (Ambion by Life Technologies, Carlsbad, CA, USA), and cDNA was synthesized by amfiRivert cDNA synthesis platinum master mix (GenDEPOT, Katy, TX, USA, Cat number R5600-200). Real-time PCR (qRT–PCR) was conducted using a 7500 FastDX (Applied Biosystems, MA, USA) and the Power SYBR green PCR master mix (Applied biosystem, Warrington WA1 4SR, UK, Cat number 4367659). Primer IDs and sequences are shown in Table S3.
Aziz F., Xin M., Gao Y., Chakroborty A., Khan I., Monts J., Monson K., Bode A.M, & Dong Z. (2020). Induction and Prevention of Gastric Cancer with Combined Helicobacter Pylori and Capsaicin Administration and DFMO Treatment, Respectively. Cancers, 12(4), 816.
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6

Quantification of Mitochondrial DNA

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Genomic DNA (gDNA) was extracted from HCT-116p53+/+ and HCT-116p53−/− cells by using DNAzol® Reagent (Thermo Scientific) and quantified using the Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (Takara Bio, Mountain View, CA, USA) according to manufacturer’s protocol. Primer sets for ND1 and ND5 were used for the detection of mtDNA, and the primer sets SLCO2B1 and SERPINA1were used for the detection of nuclear DNA. The quantitation of mtDNA copy number was performed using a 7500 FastDX (Applied Biosystems, MA, USA) and the power SYBR green PCR master mix (Applied Biosystem, Warrington WA1 4SR, UK).
De Vitto H., Ryu J., Calderon-Aparicio A., Monts J., Dey R., Chakraborty A., Lee M.H., Bode A.M, & Dong Z. (2020). Estrogen-related receptor alpha directly binds to p53 and cooperatively controls colon cancer growth through the regulation of mitochondrial biogenesis and function. Cancer & Metabolism, 8, 28.
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7

SARS-CoV-2 Qualitative Detection Using Multiple PCR Systems

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For qualitative detection of SARS-CoV-2 using both regular and Fast approaches, 3 different real time PCR systems including QuantStudio™5, Applied Biosystems™ 7500 Fast Dx, and CFX96 Touch Real-Time Detection System were used. A PCR reaction volume of 20μl containing 10μl Mastermix, 1.5μl Primer & Probe solution mix, and 8.5μl extracted RNA sample from a suspected SAR-CoV-2 patient was run on each of the mentioned thermal cyclers. Thermal cycling profile and ramp rate were optimized for each PCR system and have been elaborated as follows.
Hossain M.W., Hossain M., Arafath K., Ety S.S., Shetu M.M., Kabir M., Noor F.A, & Mannoor K. (2022). Real-Time fast PCR amplification using designated and conventional real time thermal cycler systems: COVID-19 perspective. PLoS ONE, 17(10), e0276464.
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