steponeplus system Thermo Fisher Search Results


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  • 90
    Thermo Fisher thermo fisher steponeplus
    : reverse transcription real-time polymerase chain reaction (RT-qPCR) with serial dilutions of the chimeric in vitro transcribed RNA containing both Mayaro (MAYV) and Oropouche (OROV) targets. Amplification plots for MAYV (A) and OROV (B), and linear regression for MAYV (C) and OROV (D) for ten-fold, 8-log, dilutions from 20 to 2E+08 copies, in duplicate. <t>PCR</t> efficiency in the multiplex assay was calculated with <t>StepOnePlus</t> Software v2.2 to be 98.642% [slope: -3.355, R2: 1] for MAYV, and 99.181% [slope: -3.341, R2: 1] for OROV. Ct = cycle threshold. Fluorescence values were exported to MS Excel and plotted using GraphPad Prism 6.0.
    Thermo Fisher Steponeplus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 3516 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher real time pcr qpcr steponeplus thermo fisher scientific waltham
    : reverse transcription real-time polymerase chain reaction (RT-qPCR) with serial dilutions of the chimeric in vitro transcribed RNA containing both Mayaro (MAYV) and Oropouche (OROV) targets. Amplification plots for MAYV (A) and OROV (B), and linear regression for MAYV (C) and OROV (D) for ten-fold, 8-log, dilutions from 20 to 2E+08 copies, in duplicate. <t>PCR</t> efficiency in the multiplex assay was calculated with <t>StepOnePlus</t> Software v2.2 to be 98.642% [slope: -3.355, R2: 1] for MAYV, and 99.181% [slope: -3.341, R2: 1] for OROV. Ct = cycle threshold. Fluorescence values were exported to MS Excel and plotted using GraphPad Prism 6.0.
    Real Time Pcr Qpcr Steponeplus Thermo Fisher Scientific Waltham, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher a steponeplus
    : reverse transcription real-time polymerase chain reaction (RT-qPCR) with serial dilutions of the chimeric in vitro transcribed RNA containing both Mayaro (MAYV) and Oropouche (OROV) targets. Amplification plots for MAYV (A) and OROV (B), and linear regression for MAYV (C) and OROV (D) for ten-fold, 8-log, dilutions from 20 to 2E+08 copies, in duplicate. <t>PCR</t> efficiency in the multiplex assay was calculated with <t>StepOnePlus</t> Software v2.2 to be 98.642% [slope: -3.355, R2: 1] for MAYV, and 99.181% [slope: -3.341, R2: 1] for OROV. Ct = cycle threshold. Fluorescence values were exported to MS Excel and plotted using GraphPad Prism 6.0.
    A Steponeplus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher steponeplus
    Fast qPCR assays employing SYBR Green. Genomic DNA targets were amplified as described in Methods using 0.5 ng human gDNA and 10 ng of purified wild-type or mutant Taq DNA polymerase. Reactions were cycled on the <t>StepOnePlus</t> instrument (Life Technologies) using cycling conditions consisting of 3 min at 95°C followed by 40 cycles of 3 s at 95°C, 10 s at 60°C. Genomic DNA targets are as follows: (A) 91 bp ABC, (B) 109 bp COMTE2, (C) 305 bp Numb. Sixty second extension time is required for Taq wild-type to efficiently amplify the 305 bp target (data not shown).
    Steponeplus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6958 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher steponeplus system
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Thermo Fisher abi steponeplus
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Abi Steponeplus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 384 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher steponeplus thermocycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus Thermocycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 778 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher steponeplus platform
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus Platform, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 106 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher steponeplus cycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus Cycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 301 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher steponeplus machine
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus Machine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 506 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Thermo Fisher steponeplus sequence detector
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus Sequence Detector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher steponeplus thermal cycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus Thermal Cycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 400 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi steponeplus thermocycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Abi Steponeplus Thermocycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher detection system
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 85 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher steponeplus apparatus
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus Apparatus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 89 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher steponeplus thermocyler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus Thermocyler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher abi steponeplus machine
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Abi Steponeplus Machine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi steponeplus system
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Abi Steponeplus System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher steponeplus lightcycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Steponeplus Lightcycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher abi steponeplus cycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Abi Steponeplus Cycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher applied biosystem steponeplus system
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
    Applied Biosystem Steponeplus System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher steponeplus kit
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Thermo Fisher fluorescence temperature cycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Thermo Fisher steponeplus qpcr machine
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Thermo Fisher steponeplus fast thermocycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Thermo Fisher applied biosystem steponeplus thermocycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Thermo Fisher abi steponeplus sequence detection system
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Thermo Fisher steponeplus sequence detection analyzer
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Thermo Fisher abi steponeplus platform
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Thermo Fisher abi steponeplus thermal cycler
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Thermo Fisher steponeplus abi prism platform
    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a <t>StepOnePlus</t> (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.
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    Image Search Results


    : reverse transcription real-time polymerase chain reaction (RT-qPCR) with serial dilutions of the chimeric in vitro transcribed RNA containing both Mayaro (MAYV) and Oropouche (OROV) targets. Amplification plots for MAYV (A) and OROV (B), and linear regression for MAYV (C) and OROV (D) for ten-fold, 8-log, dilutions from 20 to 2E+08 copies, in duplicate. PCR efficiency in the multiplex assay was calculated with StepOnePlus Software v2.2 to be 98.642% [slope: -3.355, R2: 1] for MAYV, and 99.181% [slope: -3.341, R2: 1] for OROV. Ct = cycle threshold. Fluorescence values were exported to MS Excel and plotted using GraphPad Prism 6.0.

    Journal: Memórias do Instituto Oswaldo Cruz

    Article Title: Multiplexed reverse transcription real-time polymerase chain reaction for simultaneous detection of Mayaro, Oropouche, and Oropouche-like viruses

    doi: 10.1590/0074-02760160062

    Figure Lengend Snippet: : reverse transcription real-time polymerase chain reaction (RT-qPCR) with serial dilutions of the chimeric in vitro transcribed RNA containing both Mayaro (MAYV) and Oropouche (OROV) targets. Amplification plots for MAYV (A) and OROV (B), and linear regression for MAYV (C) and OROV (D) for ten-fold, 8-log, dilutions from 20 to 2E+08 copies, in duplicate. PCR efficiency in the multiplex assay was calculated with StepOnePlus Software v2.2 to be 98.642% [slope: -3.355, R2: 1] for MAYV, and 99.181% [slope: -3.341, R2: 1] for OROV. Ct = cycle threshold. Fluorescence values were exported to MS Excel and plotted using GraphPad Prism 6.0.

    Article Snippet: We used the TaqMan® Fast Virus 1-Step master mix (Applied Biosystems) for RT-qPCR amplification with the recommended cycling parameters, 50ºC for 5 min for reverse transcription, 95ºC for 20 s for RT inactivation/initial denaturation, followed by 45 cycles of 95ºC for 3 s and 60ºC for 30 s. RNA (2 μL) was used as a template in a 20 µL reaction, and all assays were performed using the StepOnePlus Real-Time PCR System (Applied Biosystems).

    Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, In Vitro, Amplification, Polymerase Chain Reaction, Multiplex Assay, Software, Fluorescence, Mass Spectrometry

    Fast qPCR assays employing SYBR Green. Genomic DNA targets were amplified as described in Methods using 0.5 ng human gDNA and 10 ng of purified wild-type or mutant Taq DNA polymerase. Reactions were cycled on the StepOnePlus instrument (Life Technologies) using cycling conditions consisting of 3 min at 95°C followed by 40 cycles of 3 s at 95°C, 10 s at 60°C. Genomic DNA targets are as follows: (A) 91 bp ABC, (B) 109 bp COMTE2, (C) 305 bp Numb. Sixty second extension time is required for Taq wild-type to efficiently amplify the 305 bp target (data not shown).

    Journal: Frontiers in Microbiology

    Article Title: Compartmentalized self-replication under fast PCR cycling conditions yields Taq DNA polymerase mutants with increased DNA-binding affinity and blood resistance

    doi: 10.3389/fmicb.2014.00408

    Figure Lengend Snippet: Fast qPCR assays employing SYBR Green. Genomic DNA targets were amplified as described in Methods using 0.5 ng human gDNA and 10 ng of purified wild-type or mutant Taq DNA polymerase. Reactions were cycled on the StepOnePlus instrument (Life Technologies) using cycling conditions consisting of 3 min at 95°C followed by 40 cycles of 3 s at 95°C, 10 s at 60°C. Genomic DNA targets are as follows: (A) 91 bp ABC, (B) 109 bp COMTE2, (C) 305 bp Numb. Sixty second extension time is required for Taq wild-type to efficiently amplify the 305 bp target (data not shown).

    Article Snippet: TaqMan qPCR reactions consisted of 10 ng wild-type or mutant Taq , human genomic DNA, 0.8 mM dNTPs, 1× Taq buffer (adjusted to 95 mM KCl for Taq mutants), and 1× β-actin primer/probe (171 bp) from Life Technologies' Assays-On-Demand. qPCR reactions were run on the StepOnePlus (Life Technologies) or CFX96 (BioRad) instrument using cycling parameters indicated in the Figure legends.

    Techniques: Real-time Polymerase Chain Reaction, SYBR Green Assay, Amplification, Purification, Mutagenesis

    Fast cycling with TaqMan detection. An Assays-on-Demand assay (Life Technologies; 171 bp β-actin target) was performed using 50, 5, 0.5 ng human genomic DNA and 10 ng of purified wild-type or mutant Taq DNA polymerase. Reactions were cycled on the StepOnePlus instrument using cycling conditions consisting of 2 min at 95°C followed by 40 cycles of: (A) 10 s at 95°C, 60 s at 60°C; or (B) 10 s at 95°C, 10 s at 60°C.

    Journal: Frontiers in Microbiology

    Article Title: Compartmentalized self-replication under fast PCR cycling conditions yields Taq DNA polymerase mutants with increased DNA-binding affinity and blood resistance

    doi: 10.3389/fmicb.2014.00408

    Figure Lengend Snippet: Fast cycling with TaqMan detection. An Assays-on-Demand assay (Life Technologies; 171 bp β-actin target) was performed using 50, 5, 0.5 ng human genomic DNA and 10 ng of purified wild-type or mutant Taq DNA polymerase. Reactions were cycled on the StepOnePlus instrument using cycling conditions consisting of 2 min at 95°C followed by 40 cycles of: (A) 10 s at 95°C, 60 s at 60°C; or (B) 10 s at 95°C, 10 s at 60°C.

    Article Snippet: TaqMan qPCR reactions consisted of 10 ng wild-type or mutant Taq , human genomic DNA, 0.8 mM dNTPs, 1× Taq buffer (adjusted to 95 mM KCl for Taq mutants), and 1× β-actin primer/probe (171 bp) from Life Technologies' Assays-On-Demand. qPCR reactions were run on the StepOnePlus (Life Technologies) or CFX96 (BioRad) instrument using cycling parameters indicated in the Figure legends.

    Techniques: Purification, Mutagenesis

    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a StepOnePlus (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.

    Journal: Nucleic Acids Research

    Article Title: Correction of RT-qPCR data for genomic DNA-derived signals with ValidPrime

    doi: 10.1093/nar/gkr1259

    Figure Lengend Snippet: Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a StepOnePlus (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.

    Article Snippet: Conventional qPCR All reactions (except when indicated) were performed in duplicate 10 µl volumes using 20 ng reverse transcribed total RNA in a StepOnePlus system (Applied Biosystems) with the SsoFast EvaGreen Supermix (BioRad) and an assay concentration of 300 nM using the cycling parameters: 95°C (20 s) followed by 40 cycles at 95°C (3 s) and 60°C (20 s).

    Techniques:

    Equivalence between Cq DNA calculated by ValidPrime and RT(−) measurements. Fold ratios in linear scale (2 - Cq (RT+) /2 - Cq (RT−) ) between either the total signal (NA) measured in spiked RT(+) reactions (dark bars) or the gDNA signal (DNA) estimated by ValidPrime (VP) from RT(+) reactions (light bars) compared to the signal in RT(−) reactions. A quantity of 20 ng of cDNA from adipose tissue (hatched bars) or from kidney, were spiked with 0.30 ng gDNA to decrease the variability due to stochastic amplification observed in RT(−) reactions. Independently of the expression level of the three genes studied in RT(+) samples, the estimations by ValidPrime of the gDNA-derived signals in RT(+) were very similar to the signals measured in RT(−) reactions, as the ratio was close to 1 (illustrated by the red dashed line; mean 1.20 ± 0.29). Data are mean ± SD from two experiments in duplicate on the StepOnePlus.

    Journal: Nucleic Acids Research

    Article Title: Correction of RT-qPCR data for genomic DNA-derived signals with ValidPrime

    doi: 10.1093/nar/gkr1259

    Figure Lengend Snippet: Equivalence between Cq DNA calculated by ValidPrime and RT(−) measurements. Fold ratios in linear scale (2 - Cq (RT+) /2 - Cq (RT−) ) between either the total signal (NA) measured in spiked RT(+) reactions (dark bars) or the gDNA signal (DNA) estimated by ValidPrime (VP) from RT(+) reactions (light bars) compared to the signal in RT(−) reactions. A quantity of 20 ng of cDNA from adipose tissue (hatched bars) or from kidney, were spiked with 0.30 ng gDNA to decrease the variability due to stochastic amplification observed in RT(−) reactions. Independently of the expression level of the three genes studied in RT(+) samples, the estimations by ValidPrime of the gDNA-derived signals in RT(+) were very similar to the signals measured in RT(−) reactions, as the ratio was close to 1 (illustrated by the red dashed line; mean 1.20 ± 0.29). Data are mean ± SD from two experiments in duplicate on the StepOnePlus.

    Article Snippet: Conventional qPCR All reactions (except when indicated) were performed in duplicate 10 µl volumes using 20 ng reverse transcribed total RNA in a StepOnePlus system (Applied Biosystems) with the SsoFast EvaGreen Supermix (BioRad) and an assay concentration of 300 nM using the cycling parameters: 95°C (20 s) followed by 40 cycles at 95°C (3 s) and 60°C (20 s).

    Techniques: Amplification, Expressing, Derivative Assay

    Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a StepOnePlus (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.

    Journal: Nucleic Acids Research

    Article Title: Correction of RT-qPCR data for genomic DNA-derived signals with ValidPrime

    doi: 10.1093/nar/gkr1259

    Figure Lengend Snippet: Correction of exogenous (spiked) gDNA with ValidPrime. The data are presented in linear scale as fold ratio (2 - Cq /2 - Cq ref ), where Cq ref is the Cq NA measured on non-spiked controls and Cq refers to Cq RNA (light bars) or Cq NA (dark bars) depending on whether or not ValidPrime correction was applied (VP − /VP + ). The data are grouped based on the impact of exogenous DNA, expressed as percentage of the total signal (%DNA) in each sample. Data were collected with either 17 GOI assays on a StepOnePlus (Applied Biosystems) using mVPA1 and mVPA5 ( A ), or with 19 assays on a BioMark (Fluidigm) using mVPA1 ( B ). All assays passed the high confidence ValidPrime criteria ( Supplementary Figure S3 ). Data are presented as the mean ± SD, with ( n ) designating the number of samples in each group. cDNAs were from mouse kidney or liver for the StepOnePlus studies and mouse uterus for the BioMark study.

    Article Snippet: Specific target amplification Pre-amplification of cDNA (produced from 25 to 65 ng of total RNA) was performed in the StepOnePlus cycler (Applied Biosystems) [at 95°C for 10 min activation step followed by 14 cycles: 95°C, (15 s), 60°C, (4 min)] in a total volume of 5 µl in the presence of all primers at a concentration of 50 nM.

    Techniques:

    Equivalence between Cq DNA calculated by ValidPrime and RT(−) measurements. Fold ratios in linear scale (2 - Cq (RT+) /2 - Cq (RT−) ) between either the total signal (NA) measured in spiked RT(+) reactions (dark bars) or the gDNA signal (DNA) estimated by ValidPrime (VP) from RT(+) reactions (light bars) compared to the signal in RT(−) reactions. A quantity of 20 ng of cDNA from adipose tissue (hatched bars) or from kidney, were spiked with 0.30 ng gDNA to decrease the variability due to stochastic amplification observed in RT(−) reactions. Independently of the expression level of the three genes studied in RT(+) samples, the estimations by ValidPrime of the gDNA-derived signals in RT(+) were very similar to the signals measured in RT(−) reactions, as the ratio was close to 1 (illustrated by the red dashed line; mean 1.20 ± 0.29). Data are mean ± SD from two experiments in duplicate on the StepOnePlus.

    Journal: Nucleic Acids Research

    Article Title: Correction of RT-qPCR data for genomic DNA-derived signals with ValidPrime

    doi: 10.1093/nar/gkr1259

    Figure Lengend Snippet: Equivalence between Cq DNA calculated by ValidPrime and RT(−) measurements. Fold ratios in linear scale (2 - Cq (RT+) /2 - Cq (RT−) ) between either the total signal (NA) measured in spiked RT(+) reactions (dark bars) or the gDNA signal (DNA) estimated by ValidPrime (VP) from RT(+) reactions (light bars) compared to the signal in RT(−) reactions. A quantity of 20 ng of cDNA from adipose tissue (hatched bars) or from kidney, were spiked with 0.30 ng gDNA to decrease the variability due to stochastic amplification observed in RT(−) reactions. Independently of the expression level of the three genes studied in RT(+) samples, the estimations by ValidPrime of the gDNA-derived signals in RT(+) were very similar to the signals measured in RT(−) reactions, as the ratio was close to 1 (illustrated by the red dashed line; mean 1.20 ± 0.29). Data are mean ± SD from two experiments in duplicate on the StepOnePlus.

    Article Snippet: Specific target amplification Pre-amplification of cDNA (produced from 25 to 65 ng of total RNA) was performed in the StepOnePlus cycler (Applied Biosystems) [at 95°C for 10 min activation step followed by 14 cycles: 95°C, (15 s), 60°C, (4 min)] in a total volume of 5 µl in the presence of all primers at a concentration of 50 nM.

    Techniques: Amplification, Expressing, Derivative Assay