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    Thermo Fisher thermo fisher scietific
    ZnT3 efficiency plot . Quadruplicate ZnT3 qPCR analyses were carried out using the indicated dilutions of a cDNA pool, derived as described in the Methods section. Mean Ct values ± SD are presented for each dilution where the assay signal rose above the detection threshold. <t>PCR</t> reaction efficiency for the ZnT3 <t>TaqMan</t> assay (85%) was calculated from the slope of the line as described in the Methods section.
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    Thermo Fisher „ taqman universal pcr master mix
    ZnT3 efficiency plot . Quadruplicate ZnT3 qPCR analyses were carried out using the indicated dilutions of a cDNA pool, derived as described in the Methods section. Mean Ct values ± SD are presented for each dilution where the assay signal rose above the detection threshold. <t>PCR</t> reaction efficiency for the ZnT3 <t>TaqMan</t> assay (85%) was calculated from the slope of the line as described in the Methods section.
    „ Taqman Universal Pcr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/„ taqman universal pcr master mix/product/Thermo Fisher
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    „ taqman universal pcr master mix - by Bioz Stars, 2020-08
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    Thermo Fisher fast universal pcr master mix
    ZnT3 efficiency plot . Quadruplicate ZnT3 qPCR analyses were carried out using the indicated dilutions of a cDNA pool, derived as described in the Methods section. Mean Ct values ± SD are presented for each dilution where the assay signal rose above the detection threshold. <t>PCR</t> reaction efficiency for the ZnT3 <t>TaqMan</t> assay (85%) was calculated from the slope of the line as described in the Methods section.
    Fast Universal Pcr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 175 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 175 article reviews
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    ZnT3 efficiency plot . Quadruplicate ZnT3 qPCR analyses were carried out using the indicated dilutions of a cDNA pool, derived as described in the Methods section. Mean Ct values ± SD are presented for each dilution where the assay signal rose above the detection threshold. PCR reaction efficiency for the ZnT3 TaqMan assay (85%) was calculated from the slope of the line as described in the Methods section.

    Journal: Molecular Neurodegeneration

    Article Title: ZnT3 mRNA levels are reduced in Alzheimer's disease post-mortem brain

    doi: 10.1186/1750-1326-4-53

    Figure Lengend Snippet: ZnT3 efficiency plot . Quadruplicate ZnT3 qPCR analyses were carried out using the indicated dilutions of a cDNA pool, derived as described in the Methods section. Mean Ct values ± SD are presented for each dilution where the assay signal rose above the detection threshold. PCR reaction efficiency for the ZnT3 TaqMan assay (85%) was calculated from the slope of the line as described in the Methods section.

    Article Snippet: Each reaction contained 2 μl cDNA (corresponding to 20 ng reverse transcribed RNA), 0.25 μl TaqMan® gene expression assay for the relevant gene, 2.5 μl TaqMan® universal PCR master mix (Applied Biosystems) and 0.25 μl RNAse free water in a total volume of 5 μl.

    Techniques: Real-time Polymerase Chain Reaction, Derivative Assay, Polymerase Chain Reaction, TaqMan Assay

    Estimation of linear range of phage particles by phage PCR. (a) Real-time PCR amplification of 10-fold serial dilutions of phage particles in water. Five μl of each preparation was mixed with 15 μl of PCR pre mix containing primers specific to the arabinose promoter of the phagemid vector and the 5’-FAM probe. (b) Standard curve of the phage PCR obtained by plotting the threshold Ct value of each dilution against the log number of phage particles added to the PCR pre mix. Each data point refers to an average of two replicates (c) Titration of 3-PBA phage particles forming the antibody-alalyte-phage complex in the PHAIA plate. The phage particles were recovered from PHAIA wells incubated with 0, 1, 5, and 25 ng/ml of 3-PBA. Each column represents the mean value of three replicates and the error bar the standard deviation.

    Journal: Analytical chemistry

    Article Title: Noncompetitive Phage Anti-Immunocomplex Real-Time PCR (PHAIA-PCR) for Sensitive Detection of Small Molecules

    doi: 10.1021/ac102353z

    Figure Lengend Snippet: Estimation of linear range of phage particles by phage PCR. (a) Real-time PCR amplification of 10-fold serial dilutions of phage particles in water. Five μl of each preparation was mixed with 15 μl of PCR pre mix containing primers specific to the arabinose promoter of the phagemid vector and the 5’-FAM probe. (b) Standard curve of the phage PCR obtained by plotting the threshold Ct value of each dilution against the log number of phage particles added to the PCR pre mix. Each data point refers to an average of two replicates (c) Titration of 3-PBA phage particles forming the antibody-alalyte-phage complex in the PHAIA plate. The phage particles were recovered from PHAIA wells incubated with 0, 1, 5, and 25 ng/ml of 3-PBA. Each column represents the mean value of three replicates and the error bar the standard deviation.

    Article Snippet: TaqMan probes (5’-FAM and 5’-VIC), 7500 Fast RT-PCR system, and PCR master mix (TaqMan® Universal PCR Master Mix (2X), No Amperase® UNG) were obtained from Applied Biosystems (Carlsbad, CA).

    Techniques: Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Amplification, Plasmid Preparation, Titration, Incubation, Standard Deviation

    PHAIA-PCR and conventional PHAIA for 3-PBA and molinate. Five μl of phage particles eluted after incubation at various concentrations of analytes were mixed with 15 μl of PCR pre mix containing primers and 5’-FAM probe designed for universal amplification. Assay conditions of the PHAIA-PCR for each analyte are the same as the PHAIAs. Each value represents the mean value of three replicates (a) Amplification of 3-PBA phage. (b) Dose-response curves for 3-PBA. (c) Amplification of molinate phage. (d) Dose-response curves for molinate.

    Journal: Analytical chemistry

    Article Title: Noncompetitive Phage Anti-Immunocomplex Real-Time PCR (PHAIA-PCR) for Sensitive Detection of Small Molecules

    doi: 10.1021/ac102353z

    Figure Lengend Snippet: PHAIA-PCR and conventional PHAIA for 3-PBA and molinate. Five μl of phage particles eluted after incubation at various concentrations of analytes were mixed with 15 μl of PCR pre mix containing primers and 5’-FAM probe designed for universal amplification. Assay conditions of the PHAIA-PCR for each analyte are the same as the PHAIAs. Each value represents the mean value of three replicates (a) Amplification of 3-PBA phage. (b) Dose-response curves for 3-PBA. (c) Amplification of molinate phage. (d) Dose-response curves for molinate.

    Article Snippet: TaqMan probes (5’-FAM and 5’-VIC), 7500 Fast RT-PCR system, and PCR master mix (TaqMan® Universal PCR Master Mix (2X), No Amperase® UNG) were obtained from Applied Biosystems (Carlsbad, CA).

    Techniques: Polymerase Chain Reaction, Incubation, Amplification

    PHAIA-PCR using the 3-PBA peptide specific probe. Primers and the 5’-VIC probe designed for specific amplification of the DNA sequence encoding the anti 3-PBA/PAb 294 immunocomplex were used for 3-PBA (A) and molinate (B) PHAIA-PCR amplification. The insert in figure A plots dose-response curve as the Ct threshold values versus the 3-PBA concentration, each point represents the mean value of three replicates.

    Journal: Analytical chemistry

    Article Title: Noncompetitive Phage Anti-Immunocomplex Real-Time PCR (PHAIA-PCR) for Sensitive Detection of Small Molecules

    doi: 10.1021/ac102353z

    Figure Lengend Snippet: PHAIA-PCR using the 3-PBA peptide specific probe. Primers and the 5’-VIC probe designed for specific amplification of the DNA sequence encoding the anti 3-PBA/PAb 294 immunocomplex were used for 3-PBA (A) and molinate (B) PHAIA-PCR amplification. The insert in figure A plots dose-response curve as the Ct threshold values versus the 3-PBA concentration, each point represents the mean value of three replicates.

    Article Snippet: TaqMan probes (5’-FAM and 5’-VIC), 7500 Fast RT-PCR system, and PCR master mix (TaqMan® Universal PCR Master Mix (2X), No Amperase® UNG) were obtained from Applied Biosystems (Carlsbad, CA).

    Techniques: Polymerase Chain Reaction, Amplification, Sequencing, Concentration Assay

    Pull-down assay using biotinylated miRNA. (A) Overview of the in vitro pull-down assay. The biotinylated double-stranded miR-19a or control random RNA was incubated in cell extract (step 1) to yield a complex of the biotinylated miRNA single strand with target mRNA and RISC (step 2). The biotinylated miRNA/target mRNA complex was incubated with streptavidin beads and pulled down (step 3). The complex was treated by DNase I and reverse-transcribed (step 4). (B) The expression of miR-19a target candidate mRNAs in HEK293 cells. PTEN , known as one of miR-19a target genes, was used as a positive control. The gene, whose sequence did not match with miR-19a seed sequences, was used as the negative control. (C) Confirmation of AGO2 protein in biotinylated miRNA/target mRNA complex by western blotting with the AGO2 antibody. Biotinylated miR-19a (lane 1), biotinylated control random RNA (lane 2), and biotin (lane 3) were used for the pull-down assay and subjected to SDS-polyacrylamide gel electrophoresis and western blotting. Total cell extract was used as the positive control (lane 4). (D) Detection of target mRNAs in biotinylated miRNA/target mRNA complex by real-time RT-PCR. The relative level of each target mRNA in the complex pulled down by using biotinylated miR-19a was compared to that of the complex pulled down by using the biotinylated control random RNA. **, p

    Journal: PLoS ONE

    Article Title: Uncovering Direct Targets of MiR-19a Involved in Lung Cancer Progression

    doi: 10.1371/journal.pone.0137887

    Figure Lengend Snippet: Pull-down assay using biotinylated miRNA. (A) Overview of the in vitro pull-down assay. The biotinylated double-stranded miR-19a or control random RNA was incubated in cell extract (step 1) to yield a complex of the biotinylated miRNA single strand with target mRNA and RISC (step 2). The biotinylated miRNA/target mRNA complex was incubated with streptavidin beads and pulled down (step 3). The complex was treated by DNase I and reverse-transcribed (step 4). (B) The expression of miR-19a target candidate mRNAs in HEK293 cells. PTEN , known as one of miR-19a target genes, was used as a positive control. The gene, whose sequence did not match with miR-19a seed sequences, was used as the negative control. (C) Confirmation of AGO2 protein in biotinylated miRNA/target mRNA complex by western blotting with the AGO2 antibody. Biotinylated miR-19a (lane 1), biotinylated control random RNA (lane 2), and biotin (lane 3) were used for the pull-down assay and subjected to SDS-polyacrylamide gel electrophoresis and western blotting. Total cell extract was used as the positive control (lane 4). (D) Detection of target mRNAs in biotinylated miRNA/target mRNA complex by real-time RT-PCR. The relative level of each target mRNA in the complex pulled down by using biotinylated miR-19a was compared to that of the complex pulled down by using the biotinylated control random RNA. **, p

    Article Snippet: The quantitative real-time PCR analysis of miR-19a was performed with a TaqMan MicroRNA Reverse Transcription Kit, TaqMan 2× Universal PCR Master Mix and TaqMan MicroRNA Assay (Life Technologies).

    Techniques: Pull Down Assay, In Vitro, Incubation, Expressing, Positive Control, Sequencing, Negative Control, Western Blot, Polyacrylamide Gel Electrophoresis, Quantitative RT-PCR

    TaqMan genotyping assays discriminate seven SNPs differentiating BoHV-1.1 and BoHV-1.2 genomes. (A) Organization of the BoHV-1 genome including two unique sequences, a long one (UL) and a short one (US). The latter is flanked by two repeated and inverted sequences (IR, internal repeat; TR, terminal repeat). Shown are the localizations of the seven point mutations detected in seven BoHV-1 genes that are targeted by TaqMan genotyping assays. The nucleotides indicate the allelic version discriminating BoHV-1.1 (in boldface type) and BoHV-1.2 at each position. (B) Endpoint reading of the fluorescence generated during PCR amplification of seven target genes discriminating BoHV-1.1 (▪) and BoHV-1.2 (□). BoHV-1.1 DNA hybridized with the VIC-labeled probes, giving rise to an intense fluorescence along the x axis during amplification. BoHV-1.2 alleles hybridized with the FAM-labeled probes, resulting in a high fluorescence signal along the y axis. Different ratios of BoHV-1.1 and -1.2 DNA (1/10, 1/1, and 10/1) were mixed together and submitted to the analysis; these samples (▵) were hybridized with both the FAM- and VIC-labeled probes, resulting in increased fluorescence signals along both axes. Dot plot data were generated from endpoint readings of the fluorescence recorded after TaqMan PCR assays were performed on total DNA quantities ranging from 1 to 20 ng.

    Journal: Journal of Virology

    Article Title: Coinfection with Two Closely Related Alphaherpesviruses Results in a Highly Diversified Recombination Mosaic Displaying Negative Genetic Interference ▿

    doi: 10.1128/JVI.02474-08

    Figure Lengend Snippet: TaqMan genotyping assays discriminate seven SNPs differentiating BoHV-1.1 and BoHV-1.2 genomes. (A) Organization of the BoHV-1 genome including two unique sequences, a long one (UL) and a short one (US). The latter is flanked by two repeated and inverted sequences (IR, internal repeat; TR, terminal repeat). Shown are the localizations of the seven point mutations detected in seven BoHV-1 genes that are targeted by TaqMan genotyping assays. The nucleotides indicate the allelic version discriminating BoHV-1.1 (in boldface type) and BoHV-1.2 at each position. (B) Endpoint reading of the fluorescence generated during PCR amplification of seven target genes discriminating BoHV-1.1 (▪) and BoHV-1.2 (□). BoHV-1.1 DNA hybridized with the VIC-labeled probes, giving rise to an intense fluorescence along the x axis during amplification. BoHV-1.2 alleles hybridized with the FAM-labeled probes, resulting in a high fluorescence signal along the y axis. Different ratios of BoHV-1.1 and -1.2 DNA (1/10, 1/1, and 10/1) were mixed together and submitted to the analysis; these samples (▵) were hybridized with both the FAM- and VIC-labeled probes, resulting in increased fluorescence signals along both axes. Dot plot data were generated from endpoint readings of the fluorescence recorded after TaqMan PCR assays were performed on total DNA quantities ranging from 1 to 20 ng.

    Article Snippet: PCRs were carried out using 10 ng of dried cellular/viral DNA in a final volume of 5 μl composed of 2.5 μl of TaqMan PCR master mix (Applied Biosystems), 0.125 μl of one SNP genotyping assay mixture containing the two primers and the two probes designed for one target SNP, and 2.375 μl of nuclease-free water.

    Techniques: Fluorescence, Generated, Polymerase Chain Reaction, Amplification, Labeling

    Amplification of Epstein-Barr virus (EBV) from DNA isolated from breast cancer and blood specimens. Real time PCR using minor groove binding (MGB)-TaqMan technology was used to quantify the viral load contained in the samples. Internal repeat region (IR)1 target sequences showed that this probe amplified its respective target over a broad range and detected low levels (2.3 EBV genomes per reaction; unpublished observation). (a) A characteristic amplification plot showing the change in fluorescence (ΔRn) as a function of amplification cycle. The horizontal red line indicates the fluorescence at 10× the standard deviation of the control. The upper left arrow indicates the fluorescence detected from Daudi, an EBV-associated endemic Burkitt lymphoma. The lower arrow indicates the fluorescence of a negative control (water). The amplification, in triplicate, of the DNA from each of the patient tumor samples is indicated. (b) The standard was constructed to contain from 2 to 200,000 copies of EBV genome. The graph shows the linear regression of the Cts (the PCR cycle number when the amplification fluorescence value reaches and exceeds the predetermined background threshold value) using each of the standards. This characteristic standard line had an r 2 = 0.995 with a slope of -3.2.

    Journal: Breast Cancer Research

    Article Title: Analysis of Epstein-Barr virus reservoirs in paired blood and breast cancer primary biopsy specimens by real time PCR

    doi: 10.1186/bcr1627

    Figure Lengend Snippet: Amplification of Epstein-Barr virus (EBV) from DNA isolated from breast cancer and blood specimens. Real time PCR using minor groove binding (MGB)-TaqMan technology was used to quantify the viral load contained in the samples. Internal repeat region (IR)1 target sequences showed that this probe amplified its respective target over a broad range and detected low levels (2.3 EBV genomes per reaction; unpublished observation). (a) A characteristic amplification plot showing the change in fluorescence (ΔRn) as a function of amplification cycle. The horizontal red line indicates the fluorescence at 10× the standard deviation of the control. The upper left arrow indicates the fluorescence detected from Daudi, an EBV-associated endemic Burkitt lymphoma. The lower arrow indicates the fluorescence of a negative control (water). The amplification, in triplicate, of the DNA from each of the patient tumor samples is indicated. (b) The standard was constructed to contain from 2 to 200,000 copies of EBV genome. The graph shows the linear regression of the Cts (the PCR cycle number when the amplification fluorescence value reaches and exceeds the predetermined background threshold value) using each of the standards. This characteristic standard line had an r 2 = 0.995 with a slope of -3.2.

    Article Snippet: Amplification was performed in a total of 20 μl containing 2× Universal TaqMan PCR master mix (Applied Biosystems, Foster City, CA, USA) 900 nM of BAMHIW (forward primer, 5'-CCC AAC ACT CCA CCA CAC C-3'; reverse primers, 5'-TCT TAG GAG CTG TCC GAG GG-3'), 250 nM of Probe LIR-1MGB2 (FAM-ACACTACACACACCCACC-MGBNFQ) and 1.75 μl of water.

    Techniques: Amplification, Isolation, Real-time Polymerase Chain Reaction, Binding Assay, Fluorescence, Standard Deviation, Negative Control, Construct, Polymerase Chain Reaction