singleplex  (Thermo Fisher)


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    Structured Review

    Thermo Fisher singleplex
    The shift from extract-based to molecule-based allergy diagnosis. For molecule-based <t>singleplex</t> approaches, the number of tests to be performed can be very high. Allergen microarrays offer the advantage of testing a large panel of molecules in one single test.
    Singleplex, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/singleplex/product/Thermo Fisher
    Average 92 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    singleplex - by Bioz Stars, 2020-04
    92/100 stars

    Images

    1) Product Images from "Molecular Approach to Allergy Diagnosis and Therapy"

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy

    Journal: Yonsei Medical Journal

    doi: 10.3349/ymj.2014.55.4.839

    The shift from extract-based to molecule-based allergy diagnosis. For molecule-based singleplex approaches, the number of tests to be performed can be very high. Allergen microarrays offer the advantage of testing a large panel of molecules in one single test.
    Figure Legend Snippet: The shift from extract-based to molecule-based allergy diagnosis. For molecule-based singleplex approaches, the number of tests to be performed can be very high. Allergen microarrays offer the advantage of testing a large panel of molecules in one single test.

    Techniques Used:

    2) Product Images from "Structure-Function Investigation of Vsp Serotypes of the Spirochete Borrelia hermsii"

    Article Title: Structure-Function Investigation of Vsp Serotypes of the Spirochete Borrelia hermsii

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0007597

    B. hermsii vsp gene copy number in brain and blood. The ability of serotypes 2, 3, 13, and 27 of B. hermsii to disseminate to the brain was examined by measuring the relative vsp gene copy number by TaqMan PCR in brain and plasma of SCID mice 2 weeks after intraperitoneal inoculation with a mixed relapse serotype population (see methods for details). Results were analyzed using the ΔΔCt method adjusting for the amount of input DNA using the mouse chromosomal gene for 18S rRNA and graphed as the log 2 negative number (2 −ΔΔCt ) so that higher values present higher gene copy number relative to the borrelial chromosomal gene for 16S rRNA . Results shown for plasma represent multiplex and for brain singleplex because the sensitivity for detection of borrelial genes in the brain is 1.5–3.6 fold lower with multiplex due to the abundance of host DNA. There was significant variability in gene copy number in brain (panel A, p
    Figure Legend Snippet: B. hermsii vsp gene copy number in brain and blood. The ability of serotypes 2, 3, 13, and 27 of B. hermsii to disseminate to the brain was examined by measuring the relative vsp gene copy number by TaqMan PCR in brain and plasma of SCID mice 2 weeks after intraperitoneal inoculation with a mixed relapse serotype population (see methods for details). Results were analyzed using the ΔΔCt method adjusting for the amount of input DNA using the mouse chromosomal gene for 18S rRNA and graphed as the log 2 negative number (2 −ΔΔCt ) so that higher values present higher gene copy number relative to the borrelial chromosomal gene for 16S rRNA . Results shown for plasma represent multiplex and for brain singleplex because the sensitivity for detection of borrelial genes in the brain is 1.5–3.6 fold lower with multiplex due to the abundance of host DNA. There was significant variability in gene copy number in brain (panel A, p

    Techniques Used: Polymerase Chain Reaction, Mouse Assay, Multiplex Assay

    3) Product Images from "Development and Validation of a Multiplex, Real-Time RT PCR Assay for the Simultaneous Detection of Classical and African Swine Fever Viruses"

    Article Title: Development and Validation of a Multiplex, Real-Time RT PCR Assay for the Simultaneous Detection of Classical and African Swine Fever Viruses

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0071019

    Analytical sensitivity of RT-PCR for singleplex, duplex and triplex assays for CSFV and ASFV detection. Serial dilutions of A) in vitro transcribed CSFV RNA or B) linearised ASFV plasmid DNA were amplified in RT-PCRs containing primers and probes to detect either single, duplex or triplex targets.
    Figure Legend Snippet: Analytical sensitivity of RT-PCR for singleplex, duplex and triplex assays for CSFV and ASFV detection. Serial dilutions of A) in vitro transcribed CSFV RNA or B) linearised ASFV plasmid DNA were amplified in RT-PCRs containing primers and probes to detect either single, duplex or triplex targets.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, In Vitro, Plasmid Preparation, Amplification

    Related Articles

    Diagnostic Assay:

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy
    Article Snippet: Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations. .. Moreover, IgG interference with IgE binding might affect diagnostic outcomes.

    Clone Assay:

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy
    Article Snippet: Beginning with the cloning of the first allergens in 1988, more than 2500 allergenic molecules have been identified so far., This huge number of allergens represents one of the pitfalls of molecular allergy diagnosis, and thus, the possibility to profile patients' IgE reactivities with a large number of allergens creates the need for algorithms to distill clinically important information out of the bulk of generated data ( ). .. Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations.

    Amplification:

    Article Title: Genotyping of Brucella species using clade specific SNPs
    Article Snippet: Assays were based on a range of amplicon sizes without overlap (range 101–259 bp) and two different dye combinations were used to visualize the fragments that discriminated each SNP state (VIC/FAM, NED/PET) (Table ). .. We ran 10 mL reactions in either singleplex or multiplex with the following mastermix: 1X PCR buffer, 2 mM MgCl2 , 0.2 mM dNTPs (Invitrogen, Carlsbad, CA), 0.2 U iTaq DNA Polymerase (Bio-Rad, Hercules, CA), and molecular grade water (GIBCO, Carlsbad, CA).

    Binding Assay:

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy
    Article Snippet: MOLECULE-BASED ALLERGY DIAGNOSIS Because limitations of allergens extracts are evident, molecular allergy diagnosis has been developed as an alternative to investigate specific IgE binding to purified molecules (natural as well as recombinant). .. Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations.

    Quantitative RT-PCR:

    Article Title: Effects of the antagomiRs 15b and 200b on the altered healing pattern of diabetic mice) Effects of the antagomiRs 15b and 200b on the altered healing pattern of diabetic mice
    Article Snippet: Paragraph title: RT‐qPCR ... Reactions have been carried out in singleplex in 96‐well plates using the TaqMan Universal PCR master mix and pre‐made hydrolysis probes (Thermo Fisher Scientific).

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: Quantitative RT-PCR was run using an sds7500fast Sequence Detection System. .. Data were analyzed with Singleplex (Applied Biosystems, CA).

    Article Title: A systems biology approach reveals a link between systemic cytokines and skeletal muscle energy metabolism in a rodent smoking model and human COPD
    Article Snippet: Paragraph title: RT-qPCR validation of custom guinea pig array ... All reactions were run in singleplex on a StepOnePlus Real Time System (Applied Biosystems) at 95°C for 10 min, followed by 40 cycles at 95°C for 15 s and 60°C for 1 min. Two-fold dilution series were performed for all primer pairs to verify the linearity of the assay.

    SYBR Green Assay:

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: Real time PCR was performed using Power SYBR® Green Master Mix. .. Data were analyzed with Singleplex (Applied Biosystems, CA).

    Article Title: A systems biology approach reveals a link between systemic cytokines and skeletal muscle energy metabolism in a rodent smoking model and human COPD
    Article Snippet: The resulting cDNA was diluted 10-fold and 2.5 uL of this was used to perform qPCR in triplicate (25 uL reaction mixture volume) using the Maxima SYBR green (Thermo Scientific) and 300 nM of primers according the manufacturer’s instructions. .. All reactions were run in singleplex on a StepOnePlus Real Time System (Applied Biosystems) at 95°C for 10 min, followed by 40 cycles at 95°C for 15 s and 60°C for 1 min. Two-fold dilution series were performed for all primer pairs to verify the linearity of the assay.

    Microarray:

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy
    Article Snippet: Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations. .. Whereas in the singleplex format allergen extracts are frequently used to detect specific IgE, the current version of the microarray chip ISAC 112 offers the detection of 112 purified natural or recombinant allergens.

    Article Title: A systems biology approach reveals a link between systemic cytokines and skeletal muscle energy metabolism in a rodent smoking model and human COPD
    Article Snippet: RT-qPCR validation of custom guinea pig array Reverse transcription of 1 ug of isolated total RNA from whole-lung tissue (same RNA as were used for the microarray part) was performed using the Tetro cDNA synthesis kit (Bioline) with random hexamer primers following the manufacturer’s instructions. .. All reactions were run in singleplex on a StepOnePlus Real Time System (Applied Biosystems) at 95°C for 10 min, followed by 40 cycles at 95°C for 15 s and 60°C for 1 min. Two-fold dilution series were performed for all primer pairs to verify the linearity of the assay.

    Incubation:

    Article Title: Glucocorticoids and 11β-HSD1 are major regulators of intramyocellular protein metabolism
    Article Snippet: In a 50μL volume, 500ng of total RNA were incubated with 250μM random hexamers, 500μM dNTPs, 20U RNase inhibitor, 63U multiscribe reverse transcriptase, 5.5mM MgCl and 1× reaction buffer. .. Reactions were performed in singleplex in 10μL volumes on 96-well plates in a reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: After adding 5 μL of MMLV 5× Reaction Buffer (NEB, UK), 10mM dNTPs, 1μL ribonuclease inhibitor (NEB, UK) and 2μL of MMLV Reverse Transcriptase to reach a total volume of 25μL, the reaction mix was incubated again for 1h at 45°C. .. Data were analyzed with Singleplex (Applied Biosystems, CA).

    Article Title: 11?-HSD1 is the major regulator of the tissue-specific effects of circulating glucocorticoid excess
    Article Snippet: In a 50-μL volume, 500 ng of total RNA was incubated with 250 μM random hexamers, 500 μM dNTPs, 20 U RNase inhibitor, 63 U Multiscribe reverse transcriptase, 5.5 mM MgCl, and 1× reaction buffer. .. Reactions were performed in singleplex in 10-μL volumes on 96-well plates in reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Expressing:

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: Paragraph title: Detection of gene expression by RT-PCR ... Data were analyzed with Singleplex (Applied Biosystems, CA).

    Article Title: Plasma MicroRNA Signature Validation for Early Detection of Colorectal Cancer
    Article Snippet: Paragraph title: miRNA expression analysis ... Briefly, 2 μL of RNA were retrotranscribed and preamplified in singleplex for 12 cycles with TaqMan PreAmp Master Mix (Life Technologies).

    Real-time Polymerase Chain Reaction:

    Article Title: Glucocorticoids and 11β-HSD1 are major regulators of intramyocellular protein metabolism
    Article Snippet: Paragraph title: RNA extraction and real-time PCR ... Reactions were performed in singleplex in 10μL volumes on 96-well plates in a reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Article Title: Effects of the antagomiRs 15b and 200b on the altered healing pattern of diabetic mice) Effects of the antagomiRs 15b and 200b on the altered healing pattern of diabetic mice
    Article Snippet: One microlitre of total cDNA was used to quantify VEGF (catalogue number: mm01281449_m1), VEGFR‐2 x(catalogue number: mm01222421_m1), Ang‐1 (catalogue number: mm00456503_m1) and TK endothelial (TEK; catalogue number: mm01256894_m1) by Real‐Time qPCR, using β‐actin (catalogue number: 4352341E; Life Technologies) as the reference gene. .. Reactions have been carried out in singleplex in 96‐well plates using the TaqMan Universal PCR master mix and pre‐made hydrolysis probes (Thermo Fisher Scientific).

    Article Title: Within-host evolution of Brucella canis during a canine brucellosis outbreak in a kennel
    Article Snippet: Species identity for all isolates was confirmed by real-time PCR by a B. canis -specific assay [ ]. .. We ran these new primers in 10 μl reactions in singleplex under the following conditions: 1× PCR buffer, 2 mM MgCl2 , 0.2 mM dNTPs, 0.04 U platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA), 0.10 μM forward primer and 0.08 μM reverse primer.

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: Real time PCR was performed using Power SYBR® Green Master Mix. .. Data were analyzed with Singleplex (Applied Biosystems, CA).

    Article Title: Plasma MicroRNA Signature Validation for Early Detection of Colorectal Cancer
    Article Snippet: Briefly, 2 μL of RNA were retrotranscribed and preamplified in singleplex for 12 cycles with TaqMan PreAmp Master Mix (Life Technologies). .. The expression of each one of the target miRNAs (miR19a, miR19b, miR15b, miR29a, miR335, and miR18) , the housekeeping miR-1228, and the spike-in cel-miR-39 was evaluated with TaqMan MicroRNA Assays (Life Technologies) by real-time PCR in a Viia7 Real Time PCR system (Life Technologies).

    Article Title: 11?-HSD1 is the major regulator of the tissue-specific effects of circulating glucocorticoid excess
    Article Snippet: Paragraph title: RNA Extraction and Real-Time PCR. ... Reactions were performed in singleplex in 10-μL volumes on 96-well plates in reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Article Title: A systems biology approach reveals a link between systemic cytokines and skeletal muscle energy metabolism in a rodent smoking model and human COPD
    Article Snippet: The resulting cDNA was diluted 10-fold and 2.5 uL of this was used to perform qPCR in triplicate (25 uL reaction mixture volume) using the Maxima SYBR green (Thermo Scientific) and 300 nM of primers according the manufacturer’s instructions. .. All reactions were run in singleplex on a StepOnePlus Real Time System (Applied Biosystems) at 95°C for 10 min, followed by 40 cycles at 95°C for 15 s and 60°C for 1 min. Two-fold dilution series were performed for all primer pairs to verify the linearity of the assay.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Development and Validation of a Multiplex, Real-Time RT PCR Assay for the Simultaneous Detection of Classical and African Swine Fever Viruses
    Article Snippet: .. Optimisation of RT-PCR assays RT-PCR optimisation was performed according to the guidelines described by Agilent Technologies on artificial nucleic acid templates for CSFV or ASFV detection alongside an exogenous internal control RNA for singleplex (CSFV or ASFV), duplex (CSFV plus IC-RNA or ASFV plus IC-RNA) and triplex (CSFV, ASFV and IC-RNA) detection using the One Step SuperScript III Platinum RT-PCR kit (Invitrogen, UK). .. Concentrations of primers, probes and Mg2+ were optimised in a final reaction volume of 25 µl to obtain maximum dRn and minimal threshold cycle (CT ) The reaction mix contained 8.4 µl nucleic acid extract, 12.5 µl 2× reaction mix, a final concentration of 5 mM MgSO4 , 5 U RNAsin, 50 nM ROX and 0.5 U Superscript III reverse transcriptase/Platinum Taq mix.

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: Paragraph title: Detection of gene expression by RT-PCR ... Data were analyzed with Singleplex (Applied Biosystems, CA).

    Generated:

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy
    Article Snippet: Beginning with the cloning of the first allergens in 1988, more than 2500 allergenic molecules have been identified so far., This huge number of allergens represents one of the pitfalls of molecular allergy diagnosis, and thus, the possibility to profile patients' IgE reactivities with a large number of allergens creates the need for algorithms to distill clinically important information out of the bulk of generated data ( ). .. Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations.

    Polymerase Chain Reaction:

    Article Title: Glucocorticoids and 11β-HSD1 are major regulators of intramyocellular protein metabolism
    Article Snippet: .. Reactions were performed in singleplex in 10μL volumes on 96-well plates in a reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems). ..

    Article Title: Development and characterization of microsatellite primers in the federally endangered Astragalus bibullatus (Fabaceae) 1
    Article Snippet: All PCR reactions were three-primer reactions following . .. Loci were initially screened in singleplex using eight individuals from two natural populations, with reactions having the following final concentrations: 1× Platinum Taq Buffer, 2 mM MgCl2 , 200 nM total dNTPs, 0.15 μM fluorescently labeled primer, 0.05 μM forward primer, 0.2 μM reverse primer, and 1 unit of Platinum Taq DNA Polymerase (Invitrogen by Life Technologies, Grand Island, New York, USA).

    Article Title: Structure-Function Investigation of Vsp Serotypes of the Spirochete Borrelia hermsii
    Article Snippet: Paragraph title: DNA extraction and TaqMan PCR ... TaqMan assays were run in multiplex, with the housekeeping gene and target gene in the same well, and in singleplex, with the housekeeping gene and target gene in different wells, with pre-designed mouse 18S rRNA primers and probe (Applied Biosystems Mm4308329) to control for the amount of host input DNA.

    Article Title: Effects of the antagomiRs 15b and 200b on the altered healing pattern of diabetic mice) Effects of the antagomiRs 15b and 200b on the altered healing pattern of diabetic mice
    Article Snippet: .. Reactions have been carried out in singleplex in 96‐well plates using the TaqMan Universal PCR master mix and pre‐made hydrolysis probes (Thermo Fisher Scientific). .. The PCR reaction was monitored by using the QuantStudio 6 Flex (Thermo Fisher Scientific), and results were quantified by the 2‐ΔΔCt method for both target and reference genes.

    Article Title: Genotyping of Brucella species using clade specific SNPs
    Article Snippet: .. We ran 10 mL reactions in either singleplex or multiplex with the following mastermix: 1X PCR buffer, 2 mM MgCl2 , 0.2 mM dNTPs (Invitrogen, Carlsbad, CA), 0.2 U iTaq DNA Polymerase (Bio-Rad, Hercules, CA), and molecular grade water (GIBCO, Carlsbad, CA). .. Reactions were multiplexed in two different PCR reactions and fragments were run in a single capillary injection on an ABI 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA).

    Article Title: Within-host evolution of Brucella canis during a canine brucellosis outbreak in a kennel
    Article Snippet: .. We ran these new primers in 10 μl reactions in singleplex under the following conditions: 1× PCR buffer, 2 mM MgCl2 , 0.2 mM dNTPs, 0.04 U platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA), 0.10 μM forward primer and 0.08 μM reverse primer. .. Thermocycling parameters were an initial denaturation at 94°C for 5 min, followed by 94°C for 45 s, 65°C for 45 s and 72°C for 45 s for 35 cycles, and a final annealing step at 72°C for 5 min. VNTR PCR amplicons were discriminated through electrophoretic analysis with an ABI PRISM 3130xl automated fluorescent capillary DNA sequencer (Applied Biosystems Inc., Foster City, CA).

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: PCR primers used for the analysis (shown in Table ) were designed based on sequences deposited in the Primer 3 database: Products were run on 1% agarose gel and visualized by ethidium bromide staining. .. Data were analyzed with Singleplex (Applied Biosystems, CA).

    Article Title: Plasma MicroRNA Signature Validation for Early Detection of Colorectal Cancer
    Article Snippet: Briefly, 2 μL of RNA were retrotranscribed and preamplified in singleplex for 12 cycles with TaqMan PreAmp Master Mix (Life Technologies). .. To calculate miRNA levels, we used cycle number at threshold (Ct) values from automatic threshold, and PCR was done in triplicate.

    Article Title: 11?-HSD1 is the major regulator of the tissue-specific effects of circulating glucocorticoid excess
    Article Snippet: .. Reactions were performed in singleplex in 10-μL volumes on 96-well plates in reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems). .. Primers and probes were supplied by Applied Biosystems as premade “assay on demands.” All reactions were normalized against the housekeeping gene 18S rRNA, provided as a preoptimized control probe.

    Article Title: A systems biology approach reveals a link between systemic cytokines and skeletal muscle energy metabolism in a rodent smoking model and human COPD
    Article Snippet: All reactions were run in singleplex on a StepOnePlus Real Time System (Applied Biosystems) at 95°C for 10 min, followed by 40 cycles at 95°C for 15 s and 60°C for 1 min. Two-fold dilution series were performed for all primer pairs to verify the linearity of the assay. .. In addition, dissociation curve analysis was performed after each PCR to check for unspecific signals.

    Injection:

    Article Title: Genotyping of Brucella species using clade specific SNPs
    Article Snippet: We ran 10 mL reactions in either singleplex or multiplex with the following mastermix: 1X PCR buffer, 2 mM MgCl2 , 0.2 mM dNTPs (Invitrogen, Carlsbad, CA), 0.2 U iTaq DNA Polymerase (Bio-Rad, Hercules, CA), and molecular grade water (GIBCO, Carlsbad, CA). .. Reactions were multiplexed in two different PCR reactions and fragments were run in a single capillary injection on an ABI 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA).

    Recombinant:

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy
    Article Snippet: MOLECULE-BASED ALLERGY DIAGNOSIS Because limitations of allergens extracts are evident, molecular allergy diagnosis has been developed as an alternative to investigate specific IgE binding to purified molecules (natural as well as recombinant). .. Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations.

    DNA Extraction:

    Article Title: Structure-Function Investigation of Vsp Serotypes of the Spirochete Borrelia hermsii
    Article Snippet: Paragraph title: DNA extraction and TaqMan PCR ... TaqMan assays were run in multiplex, with the housekeeping gene and target gene in the same well, and in singleplex, with the housekeeping gene and target gene in different wells, with pre-designed mouse 18S rRNA primers and probe (Applied Biosystems Mm4308329) to control for the amount of host input DNA.

    Article Title: Within-host evolution of Brucella canis during a canine brucellosis outbreak in a kennel
    Article Snippet: Primary isolates were subcultered on trypticase soy agar plates (Difco, BD Diagnostic Systems, Sparks, MD) three times to reveal pure culture before DNA extraction (QIAamp DNA Mini Kit, Qiagen Inc., Valencia, CA). .. We ran these new primers in 10 μl reactions in singleplex under the following conditions: 1× PCR buffer, 2 mM MgCl2 , 0.2 mM dNTPs, 0.04 U platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA), 0.10 μM forward primer and 0.08 μM reverse primer.

    Isolation:

    Article Title: A systems biology approach reveals a link between systemic cytokines and skeletal muscle energy metabolism in a rodent smoking model and human COPD
    Article Snippet: RT-qPCR validation of custom guinea pig array Reverse transcription of 1 ug of isolated total RNA from whole-lung tissue (same RNA as were used for the microarray part) was performed using the Tetro cDNA synthesis kit (Bioline) with random hexamer primers following the manufacturer’s instructions. .. All reactions were run in singleplex on a StepOnePlus Real Time System (Applied Biosystems) at 95°C for 10 min, followed by 40 cycles at 95°C for 15 s and 60°C for 1 min. Two-fold dilution series were performed for all primer pairs to verify the linearity of the assay.

    Multiplex Assay:

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy
    Article Snippet: .. Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations. .. Whereas in the singleplex format allergen extracts are frequently used to detect specific IgE, the current version of the microarray chip ISAC 112 offers the detection of 112 purified natural or recombinant allergens.

    Article Title: Structure-Function Investigation of Vsp Serotypes of the Spirochete Borrelia hermsii
    Article Snippet: .. TaqMan assays were run in multiplex, with the housekeeping gene and target gene in the same well, and in singleplex, with the housekeeping gene and target gene in different wells, with pre-designed mouse 18S rRNA primers and probe (Applied Biosystems Mm4308329) to control for the amount of host input DNA. .. Custom designed primers and probes for the B. hermsii chromosomal rrs gene for 16S ribosomal RNA and the linear plasmid genes vsp3 , vsp13 , vsp2 and vsp27 were made by Applied Biosystems ( ).

    Article Title: Genotyping of Brucella species using clade specific SNPs
    Article Snippet: .. We ran 10 mL reactions in either singleplex or multiplex with the following mastermix: 1X PCR buffer, 2 mM MgCl2 , 0.2 mM dNTPs (Invitrogen, Carlsbad, CA), 0.2 U iTaq DNA Polymerase (Bio-Rad, Hercules, CA), and molecular grade water (GIBCO, Carlsbad, CA). .. Reactions were multiplexed in two different PCR reactions and fragments were run in a single capillary injection on an ABI 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA).

    Labeling:

    Article Title: Glucocorticoids and 11β-HSD1 are major regulators of intramyocellular protein metabolism
    Article Snippet: Reactions were performed in singleplex in 10μL volumes on 96-well plates in a reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems). .. All target genes were labeled with 6-carboxyfluorescein (6-FAM), and the reference gene with 2′-chloro-7′phenyl-1,4-dichloro-6-carboxy-fluorescein (VIC).

    Article Title: Development and characterization of microsatellite primers in the federally endangered Astragalus bibullatus (Fabaceae) 1
    Article Snippet: .. Loci were initially screened in singleplex using eight individuals from two natural populations, with reactions having the following final concentrations: 1× Platinum Taq Buffer, 2 mM MgCl2 , 200 nM total dNTPs, 0.15 μM fluorescently labeled primer, 0.05 μM forward primer, 0.2 μM reverse primer, and 1 unit of Platinum Taq DNA Polymerase (Invitrogen by Life Technologies, Grand Island, New York, USA). .. A touchdown cycling profile was used for all amplifications: six cycles of 95°C for 45 s, followed by annealing at 61°C (dropping one degree per cycle) for 45 s, and 72 ° C for 1 min; the remaining 29 cycles consisted of 95°C for 40 s, 55°C for 45 s, and 72°C for 1 min. All loci were initially run in singleplex on an ABI 3130xl DNA Analyzer at Middle Tennessee State University using a GeneScan 500 LIZ Size Standard (Applied Biosystems by Life Technologies, Grand Island, New York, USA).

    Article Title: Genotyping of Brucella species using clade specific SNPs
    Article Snippet: The amplification primers have unique tails that can subsequently bind to fluorescently labeled universal-tailed primers. .. We ran 10 mL reactions in either singleplex or multiplex with the following mastermix: 1X PCR buffer, 2 mM MgCl2 , 0.2 mM dNTPs (Invitrogen, Carlsbad, CA), 0.2 U iTaq DNA Polymerase (Bio-Rad, Hercules, CA), and molecular grade water (GIBCO, Carlsbad, CA).

    Purification:

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy
    Article Snippet: MOLECULE-BASED ALLERGY DIAGNOSIS Because limitations of allergens extracts are evident, molecular allergy diagnosis has been developed as an alternative to investigate specific IgE binding to purified molecules (natural as well as recombinant). .. Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations.

    Sequencing:

    Article Title: Glucocorticoids and 11β-HSD1 are major regulators of intramyocellular protein metabolism
    Article Snippet: The reverse transcription reaction was carried out at 25°C for 10min, 48°C for 30min before the reaction was terminated by heating to 95°C for 5min. mRNA levels were determined using an ABI 7500 sequence detection system (Applied Biosystems). .. Reactions were performed in singleplex in 10μL volumes on 96-well plates in a reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: Quantitative RT-PCR was run using an sds7500fast Sequence Detection System. .. Data were analyzed with Singleplex (Applied Biosystems, CA).

    Article Title: 11?-HSD1 is the major regulator of the tissue-specific effects of circulating glucocorticoid excess
    Article Snippet: The reverse transcription reaction was carried out at 25 °C for 10 min and at 48 °C for 30 min before the reaction was terminated by heating to 95 °C for 5 min. mRNA levels were determined using an ABI 7500 sequence detection system (Applied Biosystems). .. Reactions were performed in singleplex in 10-μL volumes on 96-well plates in reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Chromatin Immunoprecipitation:

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy
    Article Snippet: .. Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations. .. Whereas in the singleplex format allergen extracts are frequently used to detect specific IgE, the current version of the microarray chip ISAC 112 offers the detection of 112 purified natural or recombinant allergens.

    Plasmid Preparation:

    Article Title: Structure-Function Investigation of Vsp Serotypes of the Spirochete Borrelia hermsii
    Article Snippet: TaqMan assays were run in multiplex, with the housekeeping gene and target gene in the same well, and in singleplex, with the housekeeping gene and target gene in different wells, with pre-designed mouse 18S rRNA primers and probe (Applied Biosystems Mm4308329) to control for the amount of host input DNA. .. Custom designed primers and probes for the B. hermsii chromosomal rrs gene for 16S ribosomal RNA and the linear plasmid genes vsp3 , vsp13 , vsp2 and vsp27 were made by Applied Biosystems ( ).

    Electrophoresis:

    Article Title: Glucocorticoids and 11β-HSD1 are major regulators of intramyocellular protein metabolism
    Article Snippet: RNA integrity was assessed by electrophoresis on 1% agarose gel. .. Reactions were performed in singleplex in 10μL volumes on 96-well plates in a reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Article Title: 11?-HSD1 is the major regulator of the tissue-specific effects of circulating glucocorticoid excess
    Article Snippet: RNA integrity was assessed by electrophoresis on 1% agarose gel. .. Reactions were performed in singleplex in 10-μL volumes on 96-well plates in reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    RNA Extraction:

    Article Title: Glucocorticoids and 11β-HSD1 are major regulators of intramyocellular protein metabolism
    Article Snippet: Paragraph title: RNA extraction and real-time PCR ... Reactions were performed in singleplex in 10μL volumes on 96-well plates in a reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Article Title: 11?-HSD1 is the major regulator of the tissue-specific effects of circulating glucocorticoid excess
    Article Snippet: Paragraph title: RNA Extraction and Real-Time PCR. ... Reactions were performed in singleplex in 10-μL volumes on 96-well plates in reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Positron Emission Tomography:

    Article Title: Genotyping of Brucella species using clade specific SNPs
    Article Snippet: We used the following sequences and dyes for the universal tails: VIC-acacgcacttgacttgtcttc, FAM6-acccaactgaatagagagc, PET-ctgtccttacctcaatctc, NED-atcgactgtgttaggtcac. .. We ran 10 mL reactions in either singleplex or multiplex with the following mastermix: 1X PCR buffer, 2 mM MgCl2 , 0.2 mM dNTPs (Invitrogen, Carlsbad, CA), 0.2 U iTaq DNA Polymerase (Bio-Rad, Hercules, CA), and molecular grade water (GIBCO, Carlsbad, CA).

    Agarose Gel Electrophoresis:

    Article Title: Glucocorticoids and 11β-HSD1 are major regulators of intramyocellular protein metabolism
    Article Snippet: RNA integrity was assessed by electrophoresis on 1% agarose gel. .. Reactions were performed in singleplex in 10μL volumes on 96-well plates in a reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: PCR primers used for the analysis (shown in Table ) were designed based on sequences deposited in the Primer 3 database: Products were run on 1% agarose gel and visualized by ethidium bromide staining. .. Data were analyzed with Singleplex (Applied Biosystems, CA).

    Article Title: 11?-HSD1 is the major regulator of the tissue-specific effects of circulating glucocorticoid excess
    Article Snippet: RNA integrity was assessed by electrophoresis on 1% agarose gel. .. Reactions were performed in singleplex in 10-μL volumes on 96-well plates in reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Random Hexamer Labeling:

    Article Title: A systems biology approach reveals a link between systemic cytokines and skeletal muscle energy metabolism in a rodent smoking model and human COPD
    Article Snippet: RT-qPCR validation of custom guinea pig array Reverse transcription of 1 ug of isolated total RNA from whole-lung tissue (same RNA as were used for the microarray part) was performed using the Tetro cDNA synthesis kit (Bioline) with random hexamer primers following the manufacturer’s instructions. .. All reactions were run in singleplex on a StepOnePlus Real Time System (Applied Biosystems) at 95°C for 10 min, followed by 40 cycles at 95°C for 15 s and 60°C for 1 min. Two-fold dilution series were performed for all primer pairs to verify the linearity of the assay.

    Spectrophotometry:

    Article Title: Effects of the antagomiRs 15b and 200b on the altered healing pattern of diabetic mice) Effects of the antagomiRs 15b and 200b on the altered healing pattern of diabetic mice
    Article Snippet: Total RNA was extracted from skin samples (70 mg) using Trizol reagent (Thermo Fisher Scientific, Waltham, MA, USA), following the manufacturer's protocol, and was quantified with a spectrophotometer (NanoDrop Lite; Thermo Fisher Scientific). .. Reactions have been carried out in singleplex in 96‐well plates using the TaqMan Universal PCR master mix and pre‐made hydrolysis probes (Thermo Fisher Scientific).

    Concentration Assay:

    Article Title: Development and Validation of a Multiplex, Real-Time RT PCR Assay for the Simultaneous Detection of Classical and African Swine Fever Viruses
    Article Snippet: Optimisation of RT-PCR assays RT-PCR optimisation was performed according to the guidelines described by Agilent Technologies on artificial nucleic acid templates for CSFV or ASFV detection alongside an exogenous internal control RNA for singleplex (CSFV or ASFV), duplex (CSFV plus IC-RNA or ASFV plus IC-RNA) and triplex (CSFV, ASFV and IC-RNA) detection using the One Step SuperScript III Platinum RT-PCR kit (Invitrogen, UK). .. Concentrations of primers, probes and Mg2+ were optimised in a final reaction volume of 25 µl to obtain maximum dRn and minimal threshold cycle (CT ) The reaction mix contained 8.4 µl nucleic acid extract, 12.5 µl 2× reaction mix, a final concentration of 5 mM MgSO4 , 5 U RNAsin, 50 nM ROX and 0.5 U Superscript III reverse transcriptase/Platinum Taq mix.

    Article Title: 11?-HSD1 is the major regulator of the tissue-specific effects of circulating glucocorticoid excess
    Article Snippet: Concentration was determined spectrophotometrically at OD260 . .. Reactions were performed in singleplex in 10-μL volumes on 96-well plates in reaction buffer containing 2× TaqMan Universal PCR Master Mix (Applied Biosystems).

    Staining:

    Article Title: LINCRNA00273 promotes cancer metastasis and its G-Quadruplex promoter can serve as a novel target to inhibit cancer invasiveness
    Article Snippet: PCR primers used for the analysis (shown in Table ) were designed based on sequences deposited in the Primer 3 database: Products were run on 1% agarose gel and visualized by ethidium bromide staining. .. Data were analyzed with Singleplex (Applied Biosystems, CA).

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    Thermo Fisher il 1 beta human singleplex kit
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    The shift from extract-based to molecule-based allergy diagnosis. For molecule-based <t>singleplex</t> approaches, the number of tests to be performed can be very high. Allergen microarrays offer the advantage of testing a large panel of molecules in one single test.
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    Thermo Fisher taqman qpcr taqman singleplex
    Multiplex to <t>singleplex</t> <t>qPCR</t> Ct comparison. Graphics showing the comparison of the Ct values for each target (actin, P35S, P3, and TNOS) obtained from the multiplex assay (in black) and from the singleplex assay (in pink). The error bars show results from three independent experiments. ( N = 3, no significant differences between any pair of multiplex vs. singleplex Ct values for the same plasmid concentration and target, Student’s t test, P
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    Thermo Fisher singleplex qrt pcr
    Receiver operating characteristic (ROC) curves of a serum 6-miRNA signature (miR-19a-3p + miR-19b-3p + miR-335-5p + miR-29a-3p + miR-15b-5p + miR-18a-5p) adjusted by age and gender, based on <t>qRT-PCR</t> results. Illustrating the FIT value alone, the 6-miRNA signature alone, the faecal haemoglobin concentration (f(Hb)) alone and the combination of 6-miRNA signature+ f(Hb). Left: patients with advanced adenomas (AA) versus controls, middle: colorectal cancer (CRC) patients versus controls, right: patients with advanced colorectal neoplasia (CRC or AA) versus controls.
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    Image Search Results


    The shift from extract-based to molecule-based allergy diagnosis. For molecule-based singleplex approaches, the number of tests to be performed can be very high. Allergen microarrays offer the advantage of testing a large panel of molecules in one single test.

    Journal: Yonsei Medical Journal

    Article Title: Molecular Approach to Allergy Diagnosis and Therapy

    doi: 10.3349/ymj.2014.55.4.839

    Figure Lengend Snippet: The shift from extract-based to molecule-based allergy diagnosis. For molecule-based singleplex approaches, the number of tests to be performed can be very high. Allergen microarrays offer the advantage of testing a large panel of molecules in one single test.

    Article Snippet: Presently, manufacturers offer molecular allergy diagnosis in singleplex (i.e., Thermo Fisher ImmunoCAP, Siemens ImmuLite, and HyCor HyTec) or multiplex [Thermo Fisher Immuno-Solid phase Allergen Chip (ISAC)] formats; both systems have their advantages and limitations.

    Techniques:

    Multiplex to singleplex qPCR Ct comparison. Graphics showing the comparison of the Ct values for each target (actin, P35S, P3, and TNOS) obtained from the multiplex assay (in black) and from the singleplex assay (in pink). The error bars show results from three independent experiments. ( N = 3, no significant differences between any pair of multiplex vs. singleplex Ct values for the same plasmid concentration and target, Student’s t test, P

    Journal: BMC Biotechnology

    Article Title: Multiplex quantitative PCR for single-reaction genetically modified (GM) plant detection and identification of false-positive GM plants linked to Cauliflower mosaic virus (CaMV) infection

    doi: 10.1186/s12896-019-0571-1

    Figure Lengend Snippet: Multiplex to singleplex qPCR Ct comparison. Graphics showing the comparison of the Ct values for each target (actin, P35S, P3, and TNOS) obtained from the multiplex assay (in black) and from the singleplex assay (in pink). The error bars show results from three independent experiments. ( N = 3, no significant differences between any pair of multiplex vs. singleplex Ct values for the same plasmid concentration and target, Student’s t test, P

    Article Snippet: TaqMan qPCR TaqMan singleplex and multiplex qPCR runs were performed on a QuantStudio 6 Flex instrument (Thermo Fisher Scientific) equipped with a 384-well block.

    Techniques: Multiplex Assay, Real-time Polymerase Chain Reaction, Singleplex Assay, Plasmid Preparation, Concentration Assay

    Receiver operating characteristic (ROC) curves of a serum 6-miRNA signature (miR-19a-3p + miR-19b-3p + miR-335-5p + miR-29a-3p + miR-15b-5p + miR-18a-5p) adjusted by age and gender, based on qRT-PCR results. Illustrating the FIT value alone, the 6-miRNA signature alone, the faecal haemoglobin concentration (f(Hb)) alone and the combination of 6-miRNA signature+ f(Hb). Left: patients with advanced adenomas (AA) versus controls, middle: colorectal cancer (CRC) patients versus controls, right: patients with advanced colorectal neoplasia (CRC or AA) versus controls.

    Journal: Cancers

    Article Title: Analysis of A 6-Mirna Signature in Serum from Colorectal Cancer Screening Participants as Non-Invasive Biomarkers for Advanced Adenoma and Colorectal Cancer Detection

    doi: 10.3390/cancers11101542

    Figure Lengend Snippet: Receiver operating characteristic (ROC) curves of a serum 6-miRNA signature (miR-19a-3p + miR-19b-3p + miR-335-5p + miR-29a-3p + miR-15b-5p + miR-18a-5p) adjusted by age and gender, based on qRT-PCR results. Illustrating the FIT value alone, the 6-miRNA signature alone, the faecal haemoglobin concentration (f(Hb)) alone and the combination of 6-miRNA signature+ f(Hb). Left: patients with advanced adenomas (AA) versus controls, middle: colorectal cancer (CRC) patients versus controls, right: patients with advanced colorectal neoplasia (CRC or AA) versus controls.

    Article Snippet: Analysis of miRNA Expression by Real-Time qRTPCR MiRNA expression was assessed by singleplex qRT-PCR with preamplification using TaqMan miRNA Assays (Thermo Fisher Scientific Inc.), according to the manufacturer’s protocol.

    Techniques: Quantitative RT-PCR, Concentration Assay