2015
DOI: 10.1089/cmb.2015.0023
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Comparison of Analytic Methods for Quantitative Real-Time Polymerase Chain Reaction Data

Abstract: Polymerase chain reaction (PCR) is a laboratory procedure to amplify and simultaneously quantify targeted DNA molecules, and then detect the product of the reaction at the end of all the amplification cycles. A more modern technique, real-time PCR, also known as quantitative PCR (qPCR), detects the product after each cycle of the progressing reaction by applying a specific fluorescence technique. The quantitative methods currently used to analyze qPCR data result in varying levels of estimation quality. This s… Show more

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Cited by 6 publications
(5 citation statements)
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“…This type of error causes to falsely increase the CT of the curves with low concentration of RNA or cDNA. There is a possibility of pattern destruction, which is usually due to frequent freezing and thawing of standards or use of very low concentration patterns [23] , [54] .…”
Section: Post-analytical Errorsmentioning
confidence: 99%
“…This type of error causes to falsely increase the CT of the curves with low concentration of RNA or cDNA. There is a possibility of pattern destruction, which is usually due to frequent freezing and thawing of standards or use of very low concentration patterns [23] , [54] .…”
Section: Post-analytical Errorsmentioning
confidence: 99%
“…been separated into different wells. 5 Because both the initial template copy and reagent numbers are partitioned into these sub-samples, the linearity assumption amounts to the mathematical statement that…”
Section: Theoretical Derivationmentioning
confidence: 99%
“…Ref. [5][6][7][8][9][10][11][12][13] and the references therein) have treated analysis of qPCR data as a task in mathematical modeling, wherein parameterized equations are assumed to fully describe the signal shape. While such approaches can be powerful tools for elucidating the mechanisms driving PCR reactions, they invariably introduce model-form errors, i.e.…”
Section: Introductionmentioning
confidence: 99%
“…qPCR data are usually analyzed by the fit-point (FP) method, which assumes equal amplification efficiency between samples 5 . However, anomalies in the background fluorescence at low template input, can affect the quantification [6][7][8] . An alternative method, maxRatio, was introduced to overcome these issues 9 .…”
Section: Introductionmentioning
confidence: 99%