Molecular quantification of environmental DNA using
Digital PCR using microfluidics is a new approach that allows absolute quantification of DNA molecules. In this study digital PCR was applied to environmental samples and the effect of PCR inhibitors on DNA quantification was tested. In the control experiment using λ DNA and humic acids underestimation of λ DNA at 1/4400 of the theoretical
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ABSTRACT Droplet digital polymerase chain reaction (ddPCR) is a new technology that was recently commercialized to enable the precise quantification of target nucleic acids in a sample. ddPCR measures absolute quantities by counting nucleic acid molecules encapsulated in discrete
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Evaluation of Droplet Digital Polymerase Chain Reaction (ddPCR) for the Absolute Quantification of Aspergillus species in the Human Airway. International Journal of Molecular Sciences 2020 21 (9) 3043. https //doi/10.3390/ijms Bin Li Yuanming Li Andreas Manz Wenming Wu.
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The use of both qRT-PCR and ddPCR led to the absolute quantification of the DNA copy number. The results clearly demonstrate the importance of the choice of DNA-isolation protocol which should
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Since conventional microbiological methods are rather slow there is growing interest in detection and quantification using molecular methods. The droplet digital polymerase chain reaction (ddPCR) is a relatively new PCR method for absolute and accurate quantification without external standards.
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Dec 15 2016 · Droplet digital-PCR is a powerful recent DNA quantification technique which can be used over a range of different scientific fields and permits an absolute quantification of the samples analyzed without the need for external or internal standards (Pinheiro et al. 2012 Hayden et al. 2013 Morisset et al. 2013).
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The emerging digital PCR (dPCR) can achieve absolute quantification of copy number of DNA molecules without relying on the application of CRMs 14 15 thereby providing a potential metrological
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Sep 20 2013 · Evaluation of Digital PCR for Absolute RNA Quantification 1. Stahlberg A Kubista M Pfaffl M (2004) Comparison of reverse transcriptases in gene expression analysis. Clinical 2. White RA 3rd Blainey PC Fan HC Quake SR (2009) Digital PCR provides sensitive and
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Since conventional microbiological methods are rather slow there is growing interest in detection and quantification using molecular methods. The droplet digital polymerase chain reaction (ddPCR) is a relatively new PCR method for absolute and accurate quantification without external standards.
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Sep 01 2012 · Real-time PCR has been widely used to evaluate gene abundance in natural microbial habitats. However PCR-inhibitory substances often reduce the efficiency of PCR leading to the underestimation of target gene copy numbers. Digital PCR using microfluidics is a new approach that allows absolute quantification of DNA molecules.
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Evaluation of Digital PCR for Absolute DNA Quantification Rebecca Sanders Jim F. Huggett Claire A. Bushell Simon Cowen Daniel J. Scott and Carole
Get PriceMolecular quantification of environmental DNA using
Sep 01 2012 · Real-time PCR has been widely used to evaluate gene abundance in natural microbial habitats. However PCR-inhibitory substances often reduce the efficiency of PCR leading to the underestimation of target gene copy numbers. Digital PCR using microfluidics is a new approach that allows absolute quantification of DNA molecules.
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qPCR assays for ddPCR. ddPCR proved to be a reliable means for detection and absolute quantification of control bacterial DNA with precision as low as ten spirochetes in an individual sample. Application of this method revealed the average carriage level of B. burgdorferi in infected I. scapularis nymphs to be
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Apr 11 2016 · Digital PCR (dPCR) as an enumeration-based quantification method is capable of quantifying the DNA copy number without the help of standards. However it
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An evaluation of RT-dPCR quantification sensitivity was performed using ERCC-25 and ERCC-99 assays. Based on UV estimated values sample was diluted in 0.5 Tween 20 (Sigma) to approximately 500 250 100 50 25 10 and 5 copies per panel (equivalent to 3077 1538 615 308 154 62 and 18 copies/µL respectively).
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Apr 11 2016 · Digital PCR (dPCR) as an enumeration-based quantification method is capable of quantifying the DNA copy number without the help of standards. However it
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Absolute Quantification with Digital PCR. Digital PCR (dPCR) is a dream come true for anyone who has struggled with references and standards in making quantitative PCR (qPCR or real-time PCR) measurements. Though the techniques are related—they both aim to estimate the amount of nucleic acid in a starting sample—there is one important
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May 05 2016 · Digital PCR is rapidly being adopted in the field of DNA-based food analysis. The direct absolute quantification it offers makes it an attractive technology for routine analysis of food and feed samples for their composition possible GMO content and compliance with labelling requirements. However assessing the performance of dPCR assays is not yet well established.
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Sep 07 2020 · Digital PCR is a novel technology which can detect target sequences with relatively low abundance and obtain the absolute copy numbers of the targets. We assessed the accuracy of dPCR in TB diagnosis using more than 250 specimens and for the first time we selected M.tuberculosis-specific IS1081 in addition to widely used IS6110 as the
Get PriceEvaluation of a Droplet Digital Polymerase Chain Reaction
ABSTRACT Droplet digital polymerase chain reaction (ddPCR) is a new technology that was recently commercialized to enable the precise quantification of target nucleic acids in a sample. ddPCR measures absolute quantities by counting nucleic acid molecules encapsulated in discrete
Get PriceMeasuring Digital PCR Quality Performance Parameters and
May 05 2016 · Digital PCR is rapidly being adopted in the field of DNA-based food analysis. The direct absolute quantification it offers makes it an attractive technology for routine analysis of food and feed samples for their composition possible GMO content and compliance with labelling requirements. However assessing the performance of dPCR assays is not yet well established.
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characterise dPCR capabilities for DNA quantification less work has been performed investigating similar parameters using RT-dPCR for RNA analysis. RT-dPCR measurement using three one-step
Get PriceMolecular quantification of environmental DNA using
Digital PCR using microfluidics is a new approach that allows absolute quantification of DNA molecules. In this study digital PCR was applied to environmental samples and the effect of PCR inhibitors on DNA quantification was tested. In the control experiment using λ DNA and humic acids underestimation of λ DNA at 1/4400 of the theoretical
Get PriceAccurate quantification of supercoiled DNA by digital PCR
Apr 11 2016 · Digital PCR (dPCR) as an enumeration-based quantification method is capable of quantifying the DNA copy number without the help of standards. However it
Get PriceEvaluation of Digital PCR for Absolute DNA Quantification
Evaluation of Droplet Digital Polymerase Chain Reaction (ddPCR) for the Absolute Quantification of Aspergillus species in the Human Airway. International Journal of Molecular Sciences 2020 21 (9) 3043. https //doi/10.3390/ijms Bin Li Yuanming Li Andreas Manz Wenming Wu.
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Evaluation of Digital PCR for Absolute RNA Quantification. Sanders R. Mason D.J. Foy C.A. et al. Gene expression measurements detailing mRNA quantities are widely employed in molecular biology and are increasingly important in diagnostic fields.
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An evaluation of RT-dPCR quantification sensitivity was performed using ERCC-25 and ERCC-99 assays. Based on UV estimated values sample was diluted in 0.5 Tween 20 (Sigma) to approximately 500 250 100 50 25 10 and 5 copies per panel (equivalent to 3077 1538 615 308 154 62 and 18 copies/µL respectively).
Get PriceEvaluation of a Droplet Digital Polymerase Chain Reaction
Droplet digital polymerase chain reaction (ddPCR) is a new technology that was recently commercialized to enable the precise quantification of target nucleic acids in a sample. ddPCR measures absolute quantities by counting nucleic acid molecules encapsulated in discrete volumetrically defined water-in-oil droplet partitions.
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Main focus of GENE QUANTIFICATION web page is to describe and summarize all technical aspects involved in quantitative gene expression analysis using real-time RT-PCR and competitive RT-PCR. It illustrates the usefulness of absolute and relative quantification assays in real-time PCR and real-time RT-PCR.
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Main focus of GENE QUANTIFICATION web page is to describe and summarize all technical aspects involved in quantitative gene expression analysis using real-time RT-PCR and competitive RT-PCR. It illustrates the usefulness of absolute and relative quantification assays in real-time PCR and real-time RT-PCR.
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