quantitect taqman probe rt pcr kit (Qiagen)

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QuantiTect Probe RT PCR Kit

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For one step qRT PCR using sequence specific probes for gene expression analysis Kit contents Qiagen QuantiTect Probe RT PCR Kit 200 x 50L rxns RNA Targets Sample One step RT PCR Reaction Sequence specific Probes Ideal for Real time Quantification of RNA Targets For One step qRT PCR Using Sequence specific Probes for Gene Expression Analysis Includes 3 x 1 7mL 2x Qiagen QuantiTect Probe RT PCR Master Mix 100L Qiagen QuantiTect RT Mix 2 x 2mL RNase free Water Benefits Highly sensitive detection of low copy targets Accurate quantification over several logs of template Use of any sequence specific probe on any real time cycler No need to optimize reaction and cycling conditions

Catalog Number:

204443

Price:

645

Category:

QuantiTect Probe RT PCR Kit

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Structured Review

Qiagen quantitect taqman probe rt pcr kit

quantitect taqman probe rt pcr kit - by Bioz Stars, 2020-02

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Images

1) Product Images from "Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis"

Article Title: Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis

Journal:

doi: 10.1099/mic.0.2008/022293-0

Figure Legend Snippet: Quantitative TaqMan RT-PCR

Techniques Used: Reverse Transcription Polymerase Chain Reaction

2) Product Images from "Inactivation of the Tulane Virus, a Novel Surrogate for the Human Norovirus"

Article Title: Inactivation of the Tulane Virus, a Novel Surrogate for the Human Norovirus

Journal: Journal of food protection

doi: 10.4315/0362-028X.JFP-12-361

Figure Legend Snippet: qRT-PCR

Techniques Used: Quantitative RT-PCR

3) Product Images from "Targeting the Transforming Growth Factor-? pathway inhibits human basal-like breast cancer metastasis"

Article Title: Targeting the Transforming Growth Factor-? pathway inhibits human basal-like breast cancer metastasis

Journal: Molecular Cancer

doi: 10.1186/1476-4598-9-122

Pharmacodynamic effects of TGF-β antagonists in vivo . A . Protein extracts from snap-frozen uninvolved lung tissue of mice treated with vehicle and LY2109761 and of mice treated with controls and 1D11 were prepared and subject to Western blotting using rabbit phosphoSmad2 antibody (1:1000 dilution). Whole cell lysate from SCP2TR was used as a control (Co). Treatment with both antagonists resulted in a reduction of phosphorylated Smad2 levels. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. B . RNA was extracted from snap-frozen uninvolved kidneys of mice treated with vehicle or LY2109761 using Trizol reagent (Invitrogen) and purified using RNeasy mini columns (Qiagen) according to the manufacturer's instructions. Transcript levels of CTGF and PAI-1 were assayed using the QuantiTect™ Probe RT-PCR Kit on a Mx4000 ® Multiplex Quantitative PCR System (Stratagene). Treatment with LY2109761 significantly reduced mRNA levels of both CTGF and PAI-1 relative to GAPDH mRNA. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. C . RNA was extracted from snap-frozen lungs of mice treated with vehicle, isotype control antibody or 1D11 and transcript levels determined as described above. No significant reduction in CTGF or PAI-1 mRNA levels could be detected in the 1D11-treated group. Values represent the means and SD of two independent experiments, 3 mice per group. Unpaired 2-sided t-test was used for comparisons.

Figure Legend Snippet: Pharmacodynamic effects of TGF-β antagonists in vivo . A . Protein extracts from snap-frozen uninvolved lung tissue of mice treated with vehicle and LY2109761 and of mice treated with controls and 1D11 were prepared and subject to Western blotting using rabbit phosphoSmad2 antibody (1:1000 dilution). Whole cell lysate from SCP2TR was used as a control (Co). Treatment with both antagonists resulted in a reduction of phosphorylated Smad2 levels. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. B . RNA was extracted from snap-frozen uninvolved kidneys of mice treated with vehicle or LY2109761 using Trizol reagent (Invitrogen) and purified using RNeasy mini columns (Qiagen) according to the manufacturer's instructions. Transcript levels of CTGF and PAI-1 were assayed using the QuantiTect™ Probe RT-PCR Kit on a Mx4000 ® Multiplex Quantitative PCR System (Stratagene). Treatment with LY2109761 significantly reduced mRNA levels of both CTGF and PAI-1 relative to GAPDH mRNA. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. C . RNA was extracted from snap-frozen lungs of mice treated with vehicle, isotype control antibody or 1D11 and transcript levels determined as described above. No significant reduction in CTGF or PAI-1 mRNA levels could be detected in the 1D11-treated group. Values represent the means and SD of two independent experiments, 3 mice per group. Unpaired 2-sided t-test was used for comparisons.

Techniques Used: In Vivo, Mouse Assay, Western Blot, Purification, Reverse Transcription Polymerase Chain Reaction, Multiplex Assay, Real-time Polymerase Chain Reaction

4) Product Images from "Characterization of H5N1 Influenza Virus Variants with Hemagglutinin Mutations Isolated from Patients"

Article Title: Characterization of H5N1 Influenza Virus Variants with Hemagglutinin Mutations Isolated from Patients

Journal: mBio

doi: 10.1128/mBio.00081-15

Effects of HA mutations on viral replication in primary human airway cells. Small airway epithelial cells were infected with viruses at an MOI of 0.1. The culture supernatants were harvested at 24 (A and D), 48 (B and E), and 72 (C and F) h postinfection and assayed by quantitative real-time RT-PCR to determine the amount of progeny virus RNA. The amounts of progeny virus RNA produced by the HA mutant viruses (A, B, and C) and the seasonal human viruses (D, E, and F) are expressed relative to the amount produced by EG/D1 (wt) virus. Each data point is the mean ± SD from triplicate experiments. Colors on the x axis highlight the categories of HA mutations (A, B, and C) and virus strains (D, E, and F) as indicated. *, P

Figure Legend Snippet: Effects of HA mutations on viral replication in primary human airway cells. Small airway epithelial cells were infected with viruses at an MOI of 0.1. The culture supernatants were harvested at 24 (A and D), 48 (B and E), and 72 (C and F) h postinfection and assayed by quantitative real-time RT-PCR to determine the amount of progeny virus RNA. The amounts of progeny virus RNA produced by the HA mutant viruses (A, B, and C) and the seasonal human viruses (D, E, and F) are expressed relative to the amount produced by EG/D1 (wt) virus. Each data point is the mean ± SD from triplicate experiments. Colors on the x axis highlight the categories of HA mutations (A, B, and C) and virus strains (D, E, and F) as indicated. *, P

Techniques Used: Infection, Quantitative RT-PCR, Produced, Mutagenesis

Clone Assay:

Article Title: Development of a Rapid Fluorescent Immunochromatographic Test to Detect Respiratory Syncytial Virus
Article Snippet: To determine the Ct values corresponding to the LOD of the FICT, qRT-PCR was performed using a Quantitect Probe RT-PCR Kit (QIAGEN, Hilden, Germany) and a CFX96 Real-Time PCR Detection System. .. To produce a standard for the determination of the RNA copy number, the template was cloned into pGEM-T Easy (Promega, Madison, WI, USA).

Centrifugation:

Article Title: RNA Interference Restricts Rift Valley Fever Virus in Multiple Insect Systems
Article Snippet: The suspensions were clarified by centrifugation (5,000 × g for 10 min at 4°C), and the supernatant was used for RNA extraction with a QIAamp viral RNA minikit (Qiagen), according to the manufacturer’s protocols. .. The qRT-PCR assay was performed using an ABI 7500 cycler (Applied Biosystems) and a QuantiTect Probe RT-PCR kit (Qiagen).

Amplification:

Article Title: A Novel Squirrel Respirovirus with Putative Zoonotic Potential
Article Snippet: Cycling conditions followed the QuantiTect Probe RT-PCR Kit (Qiagen) manufacturer protocol. .. For validation, RNA covering the amplification site of MRV and HRV1 was transcribed in vitro using T7-Polymerase (Thermo Fisher) as described by the manufacturer, followed by DNase digestion (TURBO DNA-free™ Kit, Thermo Fisher), and spiked into background squirrel RNA.

Article Title: Isolation of Escherichia coli Strains with AcrAB–TolC Efflux Pump-Associated Intermediate Interpretation or Resistance to Fluoroquinolone, Chloramphenicol and Aminopenicillin from Dogs Admitted to a University Veterinary Hospital
Article Snippet: .. RT-PCR amplification mixtures (20 µl ) contained purified RNA, 2× QuantiTect Probe RT-PCR Master Mix, 0.2 µl of QuantiTect RT Mix (QuantiTect Probe RT-PCR kit, Qiagen), 0.2 µ M of probe and 0.5µ M forward and reverse primers. .. The cycle conditions comprised 20-min reverse transcription at 50°C; a 15-min initial activation step at 95°C; and 45 cycles each of 55°C for 1 min and at 60°C for 30 sec in a LightCycler 480 (Roche, Mannheim, Germany).

Article Title: Hepatitis E virus genotypes and subgenotypes causing acute hepatitis, Bulgaria, 2013–2015
Article Snippet: .. Amplification was carried out on the Rotor-Gene Q 5plex Platform (QIAGEN, Hilden, Germany) by the QuantiTect Probe RT-PCR kit (QIAGEN, Hilden, Germany). .. Data were analysed by the Rotor-Gene Q software, Version 2.1.0 (Build 9).

Positive Control:

Article Title: Saffold Virus, a Human Cardiovirus, and Risk of Persistent Islet Autoantibodies in the Longitudinal Birth Cohort Study MIDIA
Article Snippet: To develop and test the PCR method and as the positive control in the SAFV screening we used plasmids pSAF404 and delPTH5, described in detail in [ ], kindly provided by Drs. Toshiki Himeda and Yoshiro Ohara of Kanazawa Medical University School of Medicine, Japan. .. All tests were performed in duplicate using the QuantiTect Probe RT-PCR Kit (Qiagen, Hilden, Germany) and carried out on an Applied Biosystems 7300 Real Time PCR system (Applied Biosystems, Foster City, CA) with the following thermal profile: 30 minutes at 50°C, 15 minutes at 95°C followed by 45 cycles of 15 seconds at 94°C and 1 minute at 60°C.

Quantitative RT-PCR:

Article Title: Saffold Virus, a Human Cardiovirus, and Risk of Persistent Islet Autoantibodies in the Longitudinal Birth Cohort Study MIDIA
Article Snippet: To test for the presence of SAFV, an in-house one-step real-time RT-PCR was developed targeting the 5´ UTR region of SAFV. .. All tests were performed in duplicate using the QuantiTect Probe RT-PCR Kit (Qiagen, Hilden, Germany) and carried out on an Applied Biosystems 7300 Real Time PCR system (Applied Biosystems, Foster City, CA) with the following thermal profile: 30 minutes at 50°C, 15 minutes at 95°C followed by 45 cycles of 15 seconds at 94°C and 1 minute at 60°C.

Article Title: Local-Scale Diversity and Between-Year “Frozen Evolution” of Avian Influenza A Viruses in Nature
Article Snippet: The extracts were screened for IAV by the RT-qPCR method of Nagy et al. . .. Subsequently, the IAV-positive specimens were tested for all nine NA subtypes (OneStep RT–PCR kit, Qiagen) and for the most common HA subtypes: H5, H7, H9 (QuantiTect Probe RT–PCR kit, Qiagen) , , and H3, H4, H6, and H11 (OneStep RT–PCR kit, Qiagen; the primer sets are available on request).

Article Title: Real-world comparison of two molecular methods for detection of respiratory viruses
Article Snippet: Paragraph title: Real-time RT-PCR testing ... Twenty-five-μL reaction mixtures containing 5 μL of specimen RNA were tested using the QuantiTect Probe RT-PCR kit (QIAGEN) on a Smart Cycler II (Cepheid).

Article Title: A Novel Squirrel Respirovirus with Putative Zoonotic Potential
Article Snippet: Paragraph title: 2.8. Real-Time Reverse Transcription-Polymerase Chain Reaction (RT-qPCR) ... Cycling conditions followed the QuantiTect Probe RT-PCR Kit (Qiagen) manufacturer protocol.

Article Title: Development of a Rapid Fluorescent Immunochromatographic Test to Detect Respiratory Syncytial Virus
Article Snippet: .. To determine the Ct values corresponding to the LOD of the FICT, qRT-PCR was performed using a Quantitect Probe RT-PCR Kit (QIAGEN, Hilden, Germany) and a CFX96 Real-Time PCR Detection System. .. To produce a standard for the determination of the RNA copy number, the template was cloned into pGEM-T Easy (Promega, Madison, WI, USA).

Article Title: RNA Interference Restricts Rift Valley Fever Virus in Multiple Insect Systems
Article Snippet: .. The qRT-PCR assay was performed using an ABI 7500 cycler (Applied Biosystems) and a QuantiTect Probe RT-PCR kit (Qiagen). .. A total of 11 bodies corresponding to infection-positive legs/wings with C T values below 30 were pooled in TRIzol reagent for RNA extraction.

Article Title: First Detection of Heartland Virus (Bunyaviridae: Phlebovirus) from Field Collected Arthropods
Article Snippet: .. Quantitative real-time RT-PCR was performed by adding 5 μL of each extracted RNA to 50 pmol of each primer and 10 pmol of probe included in HRTV primer-probe set 1 ( ) in a 50 μL total reaction using the Qiagen QuantiTect Probe RT-PCR kit (Qiagen) according to the manufacturer's instructions. .. Reactions were conducted on Bio-Rad's CFX96 Touch real-time PCR Detection System (Bio-Rad, Hercules, CA) using the following cycling conditions: 1 cycle of 50°C for 30 min, 1 cycle of 95°C for 10 min, and 45 cycles of 95°C for 15 sec and 60°C for 1 min.

Article Title: Lack of T-bet reduces monocytic interleukin-12 formation and accelerates thrombus resolution in deep vein thrombosis
Article Snippet: Paragraph title: Quantitative RT-PCR ... For RT-PCR analysis 0.125 µg of total RNA were used with the QuantiTect Probe RT-PCR kit (Qiagen, Hilden, Germany).

Article Title: Toxic Effects of the Major Components of Diesel Exhaust in Human Alveolar Basal Epithelial Cells (A549)
Article Snippet: .. Analyses of mRNA Expression Quantitative real-time polymerase chain reaction (qRT-PCR) was performed using a QuantiTect Probe RT-PCR kit (Qiagen, Hilden, Germany) on a 7900HT Fast Real-Time PCR System (Applied Biosystems, Waltham, MA, USA). .. Each RNA sample (1 μL, 300 ng) was mixed with 1 μL of primers (stock solution, 5 μM; Generi Biotech, Hradec Kralove, Czech Republic), 0.1 μL of fluorescence probe (stock solution: for NQO1, POR, AKR1C2, COX2: 10 μM, Roche, Mannheim, Germany; for CYP1A1 and CYP1B1: 1 μM, Generi Biotech), 0.1 μL RT mix, and 5 μL RT-PCR mix and incubated 30 min at 55 °C. qRT-PCR included a 15-min incubation at 95 °C and 40 cycles of 15-sencond incubation at 95 °C followed by 1-min incubation at 60 °C.

Real-time Polymerase Chain Reaction:

Article Title: Saffold Virus, a Human Cardiovirus, and Risk of Persistent Islet Autoantibodies in the Longitudinal Birth Cohort Study MIDIA
Article Snippet: .. All tests were performed in duplicate using the QuantiTect Probe RT-PCR Kit (Qiagen, Hilden, Germany) and carried out on an Applied Biosystems 7300 Real Time PCR system (Applied Biosystems, Foster City, CA) with the following thermal profile: 30 minutes at 50°C, 15 minutes at 95°C followed by 45 cycles of 15 seconds at 94°C and 1 minute at 60°C. .. To quantify the amounts of virus present a standard curve ranging from 100 to 105 copies/μl was run alongside the samples, using 2μl of plasmid RNA as template.

Article Title: XMRV: usage of receptors and potential co-receptors
Article Snippet: .. Quantitative PCR An equal amount of total RNA was used to quantify XMRV levels using QuantiTect Probe RT-PCR kit (Qiagen). .. Real-time PCR Master Mix (Quantitect Probe RT-PCR, Qiagen) was added with forward and reverse primers, probe, and cDNA or RNA in a total volume of 25 uL.

Article Title: Molecular epidemiology of genogroup II norovirus infections in acute gastroenteritis patients during 2014–2016 in Pudong New Area, Shanghai, China
Article Snippet: NoV GI and GII positive samples were screened with QuantiTect® Probe RT-PCR Kit (Qiagen, Hilden, Germany). .. Each 20 μL reaction volume consisted of 5.7 μL RNase-free water, 12.5 μL 2× QuantiTect Probe RT-PCR Master Mix, 0.3 μL 10 mM TaqMan prober, 0.3 μL RT mix, 0.6 μL–10 mM e forward and reverse primers, respectively. qPCR was performed with a LightCycler® 480II system (Roche, Switzerland) with the thermal profile of 50 °C for 30 min; 95 °C for 15 min, 40 cycles of 95 °C for 10 s, 56 °C for 1 min.

Article Title: Hepatic Endothelial CCL25 Mediates the Recruitment of CCR9+ Gut-homing Lymphocytes to the Liver in Primary Sclerosing Cholangitis
Article Snippet: Paragraph title: Real-Time PCR. ... Each reaction was performed in triplicate using QuantiTect Probe RT-PCR kit (QIAGEN) according to the manufacturer's instructions.

Article Title: First Detection of Heartland Virus (Bunyaviridae: Phlebovirus) from Field Collected Arthropods
Article Snippet: Quantitative real-time RT-PCR was performed by adding 5 μL of each extracted RNA to 50 pmol of each primer and 10 pmol of probe included in HRTV primer-probe set 1 ( ) in a 50 μL total reaction using the Qiagen QuantiTect Probe RT-PCR kit (Qiagen) according to the manufacturer's instructions. .. Reactions were conducted on Bio-Rad's CFX96 Touch real-time PCR Detection System (Bio-Rad, Hercules, CA) using the following cycling conditions: 1 cycle of 50°C for 30 min, 1 cycle of 95°C for 10 min, and 45 cycles of 95°C for 15 sec and 60°C for 1 min.

Article Title: Isolation of Escherichia coli Strains with AcrAB–TolC Efflux Pump-Associated Intermediate Interpretation or Resistance to Fluoroquinolone, Chloramphenicol and Aminopenicillin from Dogs Admitted to a University Veterinary Hospital
Article Snippet: Purified RNA (2.5 n g) was used in one-step RT and real-time PCR amplification. .. RT-PCR amplification mixtures (20 µl ) contained purified RNA, 2× QuantiTect Probe RT-PCR Master Mix, 0.2 µl of QuantiTect RT Mix (QuantiTect Probe RT-PCR kit, Qiagen), 0.2 µ M of probe and 0.5µ M forward and reverse primers.

Article Title: Hepatitis E virus genotypes and subgenotypes causing acute hepatitis, Bulgaria, 2013–2015
Article Snippet: Paragraph title: Serum HEV RNA quantification by Real-Time PCR ... Amplification was carried out on the Rotor-Gene Q 5plex Platform (QIAGEN, Hilden, Germany) by the QuantiTect Probe RT-PCR kit (QIAGEN, Hilden, Germany).

Article Title: Lack of T-bet reduces monocytic interleukin-12 formation and accelerates thrombus resolution in deep vein thrombosis
Article Snippet: RT-PCR was performed with the CFX96 Real-Time PCR Detection System (BioRad, Munich, Germany). .. For RT-PCR analysis 0.125 µg of total RNA were used with the QuantiTect Probe RT-PCR kit (Qiagen, Hilden, Germany).

Incubation:

Article Title: XMRV: usage of receptors and potential co-receptors
Article Snippet: Quantitative PCR An equal amount of total RNA was used to quantify XMRV levels using QuantiTect Probe RT-PCR kit (Qiagen). .. The mixture was incubated at 50°C for 2 min (for RNA, 20 min), at 95°C for 10 min, and then cycled at 95°C for 15 sec and 60°C for 60 sec 40 times, using the Applied Biosystems 7500 sequence detection system.

Article Title: Toxic Effects of the Major Components of Diesel Exhaust in Human Alveolar Basal Epithelial Cells (A549)
Article Snippet: Analyses of mRNA Expression Quantitative real-time polymerase chain reaction (qRT-PCR) was performed using a QuantiTect Probe RT-PCR kit (Qiagen, Hilden, Germany) on a 7900HT Fast Real-Time PCR System (Applied Biosystems, Waltham, MA, USA). .. Each RNA sample (1 μL, 300 ng) was mixed with 1 μL of primers (stock solution, 5 μM; Generi Biotech, Hradec Kralove, Czech Republic), 0.1 μL of fluorescence probe (stock solution: for NQO1, POR, AKR1C2, COX2: 10 μM, Roche, Mannheim, Germany; for CYP1A1 and CYP1B1: 1 μM, Generi Biotech), 0.1 μL RT mix, and 5 μL RT-PCR mix and incubated 30 min at 55 °C. qRT-PCR included a 15-min incubation at 95 °C and 40 cycles of 15-sencond incubation at 95 °C followed by 1-min incubation at 60 °C.

Expressing:

Article Title: Hepatic Endothelial CCL25 Mediates the Recruitment of CCR9+ Gut-homing Lymphocytes to the Liver in Primary Sclerosing Cholangitis
Article Snippet: Each reaction was performed in triplicate using QuantiTect Probe RT-PCR kit (QIAGEN) according to the manufacturer's instructions. .. Data are given as fold increase in gene expression normalized to the 18S control and compared with normal skin tissue (normalized to 1).

Article Title: Isolation of Escherichia coli Strains with AcrAB–TolC Efflux Pump-Associated Intermediate Interpretation or Resistance to Fluoroquinolone, Chloramphenicol and Aminopenicillin from Dogs Admitted to a University Veterinary Hospital
Article Snippet: Gene expression (acrA , acrB and tolC ) was estimated by quantitative reverse transcription (RT) TaqMan-PCR. .. RT-PCR amplification mixtures (20 µl ) contained purified RNA, 2× QuantiTect Probe RT-PCR Master Mix, 0.2 µl of QuantiTect RT Mix (QuantiTect Probe RT-PCR kit, Qiagen), 0.2 µ M of probe and 0.5µ M forward and reverse primers.

Article Title: Lack of T-bet reduces monocytic interleukin-12 formation and accelerates thrombus resolution in deep vein thrombosis
Article Snippet: For RT-PCR analysis 0.125 µg of total RNA were used with the QuantiTect Probe RT-PCR kit (Qiagen, Hilden, Germany). .. TaqMan Gene Expression assays were used as probe and primer sets (Applied Biosystems, Foster City, CA) for beta actin (mouse: Mm00607939_s1), TATA-box binding protein (Tbp, mouse: Mm00446973_m-1), MCP-1 (Ccl2, mouse: Mm00441242_m1), IL-4 (mouse: Mm00445259_m1), IL-10 (mouse: Mm00439614_m1), IL-12b (mouse: Mm00434174_m1), Plasminogen activator inhibitor-1 (PAI-1, mouse: Mm00435858_m1).

Modification:

Article Title: Lack of T-bet reduces monocytic interleukin-12 formation and accelerates thrombus resolution in deep vein thrombosis
Article Snippet: For isolation of venous and thrombus RNA from snap-frozen mouse veins and thrombus material were homogenized with stainless steel micro-pestles (A. Hartenstein, Würzburg, Germany) and for RNA isolation the modified guanidine isothiocyanate method of Chomczynski and Sacchi was used. .. For RT-PCR analysis 0.125 µg of total RNA were used with the QuantiTect Probe RT-PCR kit (Qiagen, Hilden, Germany).

Concentration Assay:

Article Title: Real-world comparison of two molecular methods for detection of respiratory viruses
Article Snippet: Primer and probe sequences, concentration, and annealing temperatures are listed in Table . .. Twenty-five-μL reaction mixtures containing 5 μL of specimen RNA were tested using the QuantiTect Probe RT-PCR kit (QIAGEN) on a Smart Cycler II (Cepheid).

Article Title: Isolation of Escherichia coli Strains with AcrAB–TolC Efflux Pump-Associated Intermediate Interpretation or Resistance to Fluoroquinolone, Chloramphenicol and Aminopenicillin from Dogs Admitted to a University Veterinary Hospital
Article Snippet: The concentration of RNA was determined spectrophotometrically (BioSpectrometer, Eppendorf, Hamburg, Germany). .. RT-PCR amplification mixtures (20 µl ) contained purified RNA, 2× QuantiTect Probe RT-PCR Master Mix, 0.2 µl of QuantiTect RT Mix (QuantiTect Probe RT-PCR kit, Qiagen), 0.2 µ M of probe and 0.5µ M forward and reverse primers.

Article Title: Evolutionary analysis of the Chikungunya virus epidemic in Mexico reveals intra-host mutational hotspots in the E1 protein
Article Snippet: .. The presence of CHIKV RNA was evaluated using the QuantiTect Probe RT-PCR kit (Qiagen) in a 25μL reaction using 12.5μL of 2x reverse transcription master mix, 0.25μL of QuantiTect RT mix, 0.25μL of each primer (1μM final concentration), probe 0.15μL (0.15μM final concentration), 6.6μL of water and 5μL de RNA. .. Using the Applied Biosystems 7500 Fast system (Applied Biosystems, Foster City, USA), reverse transcription was carried out at 50°C for 30 mins followed by 95°C for 15 minutes and 45 cycles of 95°C for 15 seconds and 56°C for 1 minute.

Infection:

Article Title: RNA Interference Restricts Rift Valley Fever Virus in Multiple Insect Systems
Article Snippet: The qRT-PCR assay was performed using an ABI 7500 cycler (Applied Biosystems) and a QuantiTect Probe RT-PCR kit (Qiagen). .. A total of 11 bodies corresponding to infection-positive legs/wings with C T values below 30 were pooled in TRIzol reagent for RNA extraction.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Molecular epidemiology of genogroup II norovirus infections in acute gastroenteritis patients during 2014–2016 in Pudong New Area, Shanghai, China
Article Snippet: .. NoV GI and GII positive samples were screened with QuantiTect® Probe RT-PCR Kit (Qiagen, Hilden, Germany). .. Each 20 μL reaction volume consisted of 5.7 μL RNase-free water, 12.5 μL 2× QuantiTect Probe RT-PCR Master Mix, 0.3 μL 10 mM TaqMan prober, 0.3 μL RT mix, 0.6 μL–10 mM e forward and reverse primers, respectively. qPCR was performed with a LightCycler® 480II system (Roche, Switzerland) with the thermal profile of 50 °C for 30 min; 95 °C for 15 min, 40 cycles of 95 °C for 10 s, 56 °C for 1 min.

Article Title: Hepatic Endothelial CCL25 Mediates the Recruitment of CCR9+ Gut-homing Lymphocytes to the Liver in Primary Sclerosing Cholangitis
Article Snippet: .. Each reaction was performed in triplicate using QuantiTect Probe RT-PCR kit (QIAGEN) according to the manufacturer's instructions. .. Reactions contained 400 nM of CCL25-specific 5′-CCACACCCAAGGTGTCTTTGA-3′ and 5′-GAGCACAGCCCACCCAAT-3′ primers (AltaBioscience) and 200 nM of CCL25-specific Taqman probe, 5′-FAM-ACTGCTGCCTGGCCTACCACTACCC-TAMRA-3′; Eurogentech).

Article Title: Local-Scale Diversity and Between-Year “Frozen Evolution” of Avian Influenza A Viruses in Nature
Article Snippet: .. Subsequently, the IAV-positive specimens were tested for all nine NA subtypes (OneStep RT–PCR kit, Qiagen) and for the most common HA subtypes: H5, H7, H9 (QuantiTect Probe RT–PCR kit, Qiagen) , , and H3, H4, H6, and H11 (OneStep RT–PCR kit, Qiagen; the primer sets are available on request). ..

Article Title: Real-world comparison of two molecular methods for detection of respiratory viruses
Article Snippet: .. Twenty-five-μL reaction mixtures containing 5 μL of specimen RNA were tested using the QuantiTect Probe RT-PCR kit (QIAGEN) on a Smart Cycler II (Cepheid). ..

Article Title: A Novel Squirrel Respirovirus with Putative Zoonotic Potential
Article Snippet: .. Cycling conditions followed the QuantiTect Probe RT-PCR Kit (Qiagen) manufacturer protocol. .. For validation, RNA covering the amplification site of MRV and HRV1 was transcribed in vitro using T7-Polymerase (Thermo Fisher) as described by the manufacturer, followed by DNase digestion (TURBO DNA-free™ Kit, Thermo Fisher), and spiked into background squirrel RNA.

Article Title: Lack of T-bet reduces monocytic interleukin-12 formation and accelerates thrombus resolution in deep vein thrombosis
Article Snippet: .. For RT-PCR analysis 0.125 µg of total RNA were used with the QuantiTect Probe RT-PCR kit (Qiagen, Hilden, Germany). .. TaqMan Gene Expression assays were used as probe and primer sets (Applied Biosystems, Foster City, CA) for beta actin (mouse: Mm00607939_s1), TATA-box binding protein (Tbp, mouse: Mm00446973_m-1), MCP-1 (Ccl2, mouse: Mm00441242_m1), IL-4 (mouse: Mm00445259_m1), IL-10 (mouse: Mm00439614_m1), IL-12b (mouse: Mm00434174_m1), Plasminogen activator inhibitor-1 (PAI-1, mouse: Mm00435858_m1).

Generated:

Article Title: RNA Interference Restricts Rift Valley Fever Virus in Multiple Insect Systems
Article Snippet: The qRT-PCR assay was performed using an ABI 7500 cycler (Applied Biosystems) and a QuantiTect Probe RT-PCR kit (Qiagen). .. RVFV strain MP12 has been generated through serial passage on MRC-5 cells in the presence of 5-fluorouacil, which generated mutations in all segments leading to attenuation of the virus in mammalian hosts, which may in part be linked to temperature sensitivity ( ) However, previous studies have demonstrated that there is no difference in the levels of virus replication and dissemination between MP12 and virulent variant ZH501 in Culex pipiens mosquitoes ( ).

Polymerase Chain Reaction:

Binding Assay:

Article Title: A Novel Squirrel Respirovirus with Putative Zoonotic Potential
Article Snippet: For a pan rodent respirovirus RT-qPCR (MGSqRV RT-qPCR) simultaneously detecting MRV and GSqRV, primers binding to MRV (MRV-fw 5′-CTGCTGACTCCTGTTTCAAC-3′, MRV-rv 5′-TGTTATRAACCGACTTGCACG-3′) were added. .. Cycling conditions followed the QuantiTect Probe RT-PCR Kit (Qiagen) manufacturer protocol.

Fluorescence:

Isolation:

Article Title: Local-Scale Diversity and Between-Year “Frozen Evolution” of Avian Influenza A Viruses in Nature
Article Snippet: Paragraph title: Virus detection and isolation ... Subsequently, the IAV-positive specimens were tested for all nine NA subtypes (OneStep RT–PCR kit, Qiagen) and for the most common HA subtypes: H5, H7, H9 (QuantiTect Probe RT–PCR kit, Qiagen) , , and H3, H4, H6, and H11 (OneStep RT–PCR kit, Qiagen; the primer sets are available on request).

Size-exclusion Chromatography:

Article Title: Isolation of Escherichia coli Strains with AcrAB–TolC Efflux Pump-Associated Intermediate Interpretation or Resistance to Fluoroquinolone, Chloramphenicol and Aminopenicillin from Dogs Admitted to a University Veterinary Hospital
Article Snippet: RT-PCR amplification mixtures (20 µl ) contained purified RNA, 2× QuantiTect Probe RT-PCR Master Mix, 0.2 µl of QuantiTect RT Mix (QuantiTect Probe RT-PCR kit, Qiagen), 0.2 µ M of probe and 0.5µ M forward and reverse primers. .. The cycle conditions comprised 20-min reverse transcription at 50°C; a 15-min initial activation step at 95°C; and 45 cycles each of 55°C for 1 min and at 60°C for 30 sec in a LightCycler 480 (Roche, Mannheim, Germany).

Labeling:

Article Title: Isolation of Escherichia coli Strains with AcrAB–TolC Efflux Pump-Associated Intermediate Interpretation or Resistance to Fluoroquinolone, Chloramphenicol and Aminopenicillin from Dogs Admitted to a University Veterinary Hospital
Article Snippet: The probes were labeled by the manufacturer (Sigma-Aldrich) with the reporter dye 6-carboxyfluorescein (6′-FAM) at the 5′-end and with the quencher dye 6-carboxytetramethylrhodamine (TAMRA) at the 3′-end. .. RT-PCR amplification mixtures (20 µl ) contained purified RNA, 2× QuantiTect Probe RT-PCR Master Mix, 0.2 µl of QuantiTect RT Mix (QuantiTect Probe RT-PCR kit, Qiagen), 0.2 µ M of probe and 0.5µ M forward and reverse primers.

Purification:

Sequencing:

Article Title: Local-Scale Diversity and Between-Year “Frozen Evolution” of Avian Influenza A Viruses in Nature
Article Snippet: Subsequently, the IAV-positive specimens were tested for all nine NA subtypes (OneStep RT–PCR kit, Qiagen) and for the most common HA subtypes: H5, H7, H9 (QuantiTect Probe RT–PCR kit, Qiagen) , , and H3, H4, H6, and H11 (OneStep RT–PCR kit, Qiagen; the primer sets are available on request). .. The results of conventional RT-PCR reactions were confirmed by sequencing and BLAST analysis conducted by the National Center for Biotechnology Information (NCBI).

Article Title: A Novel Squirrel Respirovirus with Putative Zoonotic Potential
Article Snippet: The full-genome sequence of GSqRV served as basis for the development of a specific RT-qPCR, amplifying a 105 nucleotide (nt) long sequence in the L gene. .. Cycling conditions followed the QuantiTect Probe RT-PCR Kit (Qiagen) manufacturer protocol.

Article Title: Isolation of Escherichia coli Strains with AcrAB–TolC Efflux Pump-Associated Intermediate Interpretation or Resistance to Fluoroquinolone, Chloramphenicol and Aminopenicillin from Dogs Admitted to a University Veterinary Hospital
Article Snippet: The respective primer pairs and probes ( ) used foracrB , tolC and gapA in this study were designed according to the sequence of E. coli strain K12 substrain MG1655, which is deposited in GenBank (accession number U00096). .. RT-PCR amplification mixtures (20 µl ) contained purified RNA, 2× QuantiTect Probe RT-PCR Master Mix, 0.2 µl of QuantiTect RT Mix (QuantiTect Probe RT-PCR kit, Qiagen), 0.2 µ M of probe and 0.5µ M forward and reverse primers.

Article Title: Hepatitis E virus genotypes and subgenotypes causing acute hepatitis, Bulgaria, 2013–2015
Article Snippet: A 70 bp region of the ORF3 (position 5261–5330 on full length HEV genome sequence Acc.N. ) was amplified from 7 μL RNA extract (equivalent to RNA extracted from 35 μL serum). .. Amplification was carried out on the Rotor-Gene Q 5plex Platform (QIAGEN, Hilden, Germany) by the QuantiTect Probe RT-PCR kit (QIAGEN, Hilden, Germany).

Plasmid Preparation:

Article Title: Saffold Virus, a Human Cardiovirus, and Risk of Persistent Islet Autoantibodies in the Longitudinal Birth Cohort Study MIDIA
Article Snippet: Plasmid RNA was synthesised by use of the RiboMAX Large Scale RNA Production Systems (Promega, Madison, WI). .. All tests were performed in duplicate using the QuantiTect Probe RT-PCR Kit (Qiagen, Hilden, Germany) and carried out on an Applied Biosystems 7300 Real Time PCR system (Applied Biosystems, Foster City, CA) with the following thermal profile: 30 minutes at 50°C, 15 minutes at 95°C followed by 45 cycles of 15 seconds at 94°C and 1 minute at 60°C.

Software:

Article Title: Hepatitis E virus genotypes and subgenotypes causing acute hepatitis, Bulgaria, 2013–2015
Article Snippet: Amplification was carried out on the Rotor-Gene Q 5plex Platform (QIAGEN, Hilden, Germany) by the QuantiTect Probe RT-PCR kit (QIAGEN, Hilden, Germany). .. Data were analysed by the Rotor-Gene Q software, Version 2.1.0 (Build 9).

RNA Extraction:

Article Title: First Detection of Heartland Virus (Bunyaviridae: Phlebovirus) from Field Collected Arthropods
Article Snippet: Paragraph title: RNA extraction and virus detection. ... Quantitative real-time RT-PCR was performed by adding 5 μL of each extracted RNA to 50 pmol of each primer and 10 pmol of probe included in HRTV primer-probe set 1 ( ) in a 50 μL total reaction using the Qiagen QuantiTect Probe RT-PCR kit (Qiagen) according to the manufacturer's instructions.

Article Title: Evolutionary analysis of the Chikungunya virus epidemic in Mexico reveals intra-host mutational hotspots in the E1 protein
Article Snippet: Viral RNA was extracted from 200μL of patient serum using the QiAmp Viral RNA Extraction Kit (Qiagen, Hilden, Germany). .. The presence of CHIKV RNA was evaluated using the QuantiTect Probe RT-PCR kit (Qiagen) in a 25μL reaction using 12.5μL of 2x reverse transcription master mix, 0.25μL of QuantiTect RT mix, 0.25μL of each primer (1μM final concentration), probe 0.15μL (0.15μM final concentration), 6.6μL of water and 5μL de RNA.

In Vitro:

Article Title: Development of a Rapid Fluorescent Immunochromatographic Test to Detect Respiratory Syncytial Virus
Article Snippet: To determine the Ct values corresponding to the LOD of the FICT, qRT-PCR was performed using a Quantitect Probe RT-PCR Kit (QIAGEN, Hilden, Germany) and a CFX96 Real-Time PCR Detection System. .. In vitro transcription reactions were performed using a RiboMax T7 transcription kit (Promega).

Sampling:

Article Title: Local-Scale Diversity and Between-Year “Frozen Evolution” of Avian Influenza A Viruses in Nature
Article Snippet: The GPS coordinates of the sampling localities were provided in . .. Subsequently, the IAV-positive specimens were tested for all nine NA subtypes (OneStep RT–PCR kit, Qiagen) and for the most common HA subtypes: H5, H7, H9 (QuantiTect Probe RT–PCR kit, Qiagen) , , and H3, H4, H6, and H11 (OneStep RT–PCR kit, Qiagen; the primer sets are available on request).

Activation Assay:

Article Title: Isolation of Escherichia coli Strains with AcrAB–TolC Efflux Pump-Associated Intermediate Interpretation or Resistance to Fluoroquinolone, Chloramphenicol and Aminopenicillin from Dogs Admitted to a University Veterinary Hospital
Article Snippet: RT-PCR amplification mixtures (20 µl ) contained purified RNA, 2× QuantiTect Probe RT-PCR Master Mix, 0.2 µl of QuantiTect RT Mix (QuantiTect Probe RT-PCR kit, Qiagen), 0.2 µ M of probe and 0.5µ M forward and reverse primers. .. The cycle conditions comprised 20-min reverse transcription at 50°C; a 15-min initial activation step at 95°C; and 45 cycles each of 55°C for 1 min and at 60°C for 30 sec in a LightCycler 480 (Roche, Mannheim, Germany).

Virus Isolation Assay:

Article Title: Local-Scale Diversity and Between-Year “Frozen Evolution” of Avian Influenza A Viruses in Nature
Article Snippet: The swabs were re-suspended in PBS buffer and the suspensions were then divided into aliquots and used either for molecular detection or virus isolation. .. Subsequently, the IAV-positive specimens were tested for all nine NA subtypes (OneStep RT–PCR kit, Qiagen) and for the most common HA subtypes: H5, H7, H9 (QuantiTect Probe RT–PCR kit, Qiagen) , , and H3, H4, H6, and H11 (OneStep RT–PCR kit, Qiagen; the primer sets are available on request).

Variant Assay:

Article Title: RNA Interference Restricts Rift Valley Fever Virus in Multiple Insect Systems
Article Snippet: The qRT-PCR assay was performed using an ABI 7500 cycler (Applied Biosystems) and a QuantiTect Probe RT-PCR kit (Qiagen). .. RVFV strain MP12 has been generated through serial passage on MRC-5 cells in the presence of 5-fluorouacil, which generated mutations in all segments leading to attenuation of the virus in mammalian hosts, which may in part be linked to temperature sensitivity ( ) However, previous studies have demonstrated that there is no difference in the levels of virus replication and dissemination between MP12 and virulent variant ZH501 in Culex pipiens mosquitoes ( ).