Abi 7500 fast dx real time pcr system

Manufactured by Thermo Fisher Scientific

Sourced in United States

The ABI 7500 Fast Dx real-time PCR system is a laboratory instrument designed for nucleic acid amplification and detection. It features a 96-well format and is capable of performing real-time quantitative PCR (qPCR) analysis.

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Lab products found in correlation

Magmax viral pathogen nucleic acid isolation kit, by Thermo Fisher Scientific (2 mentions) Kingfisher flex instrument, by Thermo Fisher Scientific (2 mentions) Taqpath 1 step multiplex master mix no rox, by Thermo Fisher Scientific (2 mentions) Nanodrop, by Thermo Fisher Scientific (2 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (1 mentions) Dneasy blood and tissue kit, by Qiagen (1 mentions) Sybr advantage qpcr premix kit, by Takara Bio (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Lightcycler 2, by Roche (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Taqman fast advanced master mix, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

ABI 7500 Real-Time PCR System (2776 citations) ABI 7500 Fast Real-Time PCR System (1243 citations) ABI 7500 Fast System (177 citations) ABI 7500 PCR system (164 citations) ABI 7500 Real-Time System (117 citations) 7500 Fast Real-Time PCR (92 citations) 7500 Real-Time PCR (79 citations) ABI 7500 Real-Time PCR (51 citations) ABI Real Time PCR System (51 citations) 7500 Fast PCR system (42 citations)

6 protocols using abi 7500 fast dx real time pcr system

SARS-CoV-2 Detection in Animal Specimens

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For animal specimens, the nucleic acid extractions were performed with the Kingfisher Flex Instrument (Thermo Fisher Inc., Waltham, MA, USA), using the MagMAX™ Viral/Pathogen Nucleic Acid Isolation Kit (Thermo Fisher Inc., Waltham, MA, USA), according to the manufacturers’ instructions. The MVP_Flex_200 ul protocol was used with 200 µL specimen volume and an elution volume of 100 µL. Human specimen control (HSC; A549 cell suspension) was included as an extraction control and water as a negative control.
rRT-PCR testing of animal specimens for SARS-CoV-2 was performed on the ABI 7500 Fast Dx Real-time PCR system (Thermo Fisher Inc., Waltham, MA, USA) using the CDC influenza SARS-CoV-2 (FluSC2) multiplex assay (https://www.fda.gov/media/139743/download; accessed on 1 November 2022) and TaqPath™ 1-Step Multiplex Master Mix (No ROX) (Thermo Fisher Scientific Inc., Waltham, MA, USA). rRT-PCR negative specimens were further tested for β-Actin using the Taq polymerase enzyme mentioned above. An animal was classified as “rRT-PCR positive” if specimens from the animal tested positive by at least one diagnostic specimen (oropharyngeal swab, nasal swab, or rectal swab).
Virus isolation was performed by USDA-NVSL as previously described [34 (link)].

Cossaboom C.M., Wendling N.M., Lewis N.M., Rettler H., Harvey R.R., Amman B.R., Towner J.S., Spengler J.R., Erickson R., Burnett C., Young E.L., Oakeson K., Carpenter A., Kainulainen M.H., Chatterjee P., Flint M., Uehara A., Li Y., Zhang J., Kelleher A., Lynch B., Retchless A.C., Tong S., Ahmad A., Bunkley P., Godino C., Herzegh O., Drobeniuc J., Rooney J., Taylor D, & Barton Behravesh C. (2022). One Health Investigation of SARS-CoV-2 in People and Animals on Multiple Mink Farms in Utah. Viruses, 15(1), 96.

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Molecular Screening of Mosquito Pools for W. bancrofti

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Pooled mosquitoes were tested for presence of W. bancrofti DNA at the molecular laboratory of the Institute of Epidemiology, Disease Control & Research (IEDCR, MoHFW) in Dhaka, Bangladesh. Positive and negative controls were also tested. DNA was extracted from pools using DNeasy Blood and Tissue kits (Qiagen, Hilden, Germany) following the manufacturer’s instructions. DNA quality was confirmed prior to PCR using a NanoDrop (Thermo Fisher, Waltham, MA, USA). The real-time PCR protocol described by Rao et al. [17 (link)] was used to detect W. bancrofti DNA in the pools. All reactions were carried out in an ABI 7500 Fast Dx real-time PCR system (Thermo Fisher Scientific, Waltham, MA, USA) using Taqman Universal PCR Master Mix (Thermo Fisher Scientific, Waltham, MA, USA), and all pools were run in duplicate.

Irish S.R., Al-Amin H.M., Paulin H.N., Mahmood A.S., Khan R.K., Muraduzzaman A.K., Worrell C.M., Flora M.S., Karim M.J., Shirin T., Shamsuzzaman A.K., Tahmina S., Lenhart A, & Dubray C. (2018). Molecular xenomonitoring for Wuchereria bancrofti in Culex quinquefasciatus in two districts in Bangladesh supports transmission assessment survey findings. PLoS Neglected Tropical Diseases, 12(7), e0006574.

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Quantitative RT-PCR analysis of adiponectin receptors

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Total RNA was extracted from the cells with TRIzol (Invitrogen, USA) according to the manufacturer's instructions. Template cDNA was prepared and then was amplified with qPCR primers designed and synthesized by GenePharma (Shanghai, China). Reverse transcription was performed on an ABI 7500 Fast Dx real-time PCR system (Thermo Fisher Scientific, Inc.) according to the following program: reverse transcription at 50°C for 30 min, initial denaturation at 94°C for 2 min, and then 45 cycles of 95°C for 15 s, 55°C for 1 min, and 72°C for 5 s. RT-PCR was carried out with a LightCycler 2.0 (Roche) using the SYBR® Advantage® qPCR Premix Kit (Clontech, Mountain View, CA). PCR primer sequences of AdipoR1, AdipoR2 and GAPDH, used as an internal control, were shown in Supplementary Table 2.

Liu J., Sui H., Zhao J, & Wang Y. (2017). Osmotin Protects H9c2 Cells from Simulated Ischemia-Reperfusion Injury through AdipoR1/PI3K/AKT Signaling Pathway. Frontiers in Physiology, 8, 611.

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Mosquito RNA Extraction and RT-qPCR Detection

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After thawing at room temperature, RNA was extracted from 100 µl of the mosquito lysate using the RNEasy Mini Kit (Qiagen, Hilden, Germany) and analyzed by RT-qPCR using the ZDC (Zika, Dengue and Chikungunya) Multiplex RT-PCR assay (Bio-Rad, Hercules, CA, USA) according to the manufacturer’s instructions on an ABI 7500 Fast Dx Real-Time PCR system (Applied Biosystems, Hercules, CA, USA).

Valentine M.J., Ciraola B., Aliota M.T., Vandenplas M., Marchi S., Tenebray B., Leparc-Goffart I., Gallagher C.A., Beierschmitt A., Corey T., Dore K.M., de Lamballerie X., Wang C., Murdock C.C, & Kelly P.J. (2020). No evidence for sylvatic cycles of chikungunya, dengue and Zika viruses in African green monkeys (Chlorocebus aethiops sabaeus) on St. Kitts, West Indies. Parasites & Vectors, 13, 540.

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Quantitative Real-Time PCR Assay Protocol

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All performance characteristics were determined by TaqMan real-time PCR assay using the ABI 7500 Fast Dx Real-time PCR system (Applied Biosystems Inc., Foster City, CA; USA.). Each sample was analyzed in three independent runs and in triplicate PCR reactions of 20μL. We used TaqMan^® Fast Advanced Master Mix (Cat # 4444556, ThermoFisher Scientific, Waltham, MA; USA) and followed the manufacturer’s instructions. Briefly, each PCR reaction mixture contained 1X Master Mix and 5μL of template DNA. The final concentration of each primer and probe was 500nM and 100nM respectively. Each real-time PCR run also included two positive controls (RD and L20B cell lines; 50pg/μL) and a no template control (NTC). The PCR amplification conditions were 50°C for 2 minutes (for the activation of uracil-N-glycosylase (UNG) in the TaqMan master mix) and 95°C for 2 minutes, followed by 45 cycles of 95°C for 3 seconds and 60°C for 30 seconds. We followed a strict uni-directional workflow to prevent any cross-contamination by using separate rooms for the preparations of the sample, master mix, and PCR amplification.

Secor W.E, & Freeman GL J.r. (2001). Differential Vβ T-Cell Receptor Usage during Chronic Experimental Schistosomiasis Corresponds with Distinct Pathological Presentations. Infection and Immunity, 69(6), 4177-4179.

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SARS-CoV-2 Detection Using Automated Nucleic Acid Extraction and qRT-PCR

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Nucleic acid extractions were performed with the Kingfisher Flex Instrument (ThermoFisher Inc., Waltham, MA, USA), using the MagMAX™ Viral/Pathogen Nucleic Acid Isolation Kit (Cat # A48310, ThermoFisher Inc., Waltham, MA, USA) according to the manufacturers’ instructions. The MVP_Flex_200ul protocol was used with an elution volume of 100 µL. Human specimen control (HSC; A549 cell suspension) was included as an extraction control and water as a negative control.
QRT-PCR testing of specimens targeting the N-gene of SARS-CoV-2 was performed on the ABI 7500 Fast Dx Real-time PCR system (Applied Biosystems, ThermoFisher Inc., Waltham, MA, USA) using the CDC influenza SARS-CoV-2 (FluSC2) multiplex assay (https://www.fda.gov/media/139743/download; kit Cat # Flu SC2-EUA, accessed on 29 April 2022). TaqPath™ 1-Step Multiplex Master Mix (No ROX) (Cat # A28522, ThermoFisher Scientific Inc., Waltham, MA, USA) was used to further test the qRT-PCR negative specimens for β-Actin to verify successful RNA extraction.

Amman B.R., Cossaboom C.M., Wendling N.M., Harvey R.R., Rettler H., Taylor D., Kainulainen M.H., Ahmad A., Bunkley P., Godino C., Tong S., Li Y., Uehara A., Kelleher A., Zhang J., Lynch B., Behravesh C.B, & Towner J.S. (2022). GPS Tracking of Free-Roaming Cats (Felis catus) on SARS-CoV-2-Infected Mink Farms in Utah. Viruses, 14(10), 2131.

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