Poisson Plus Quantification for Digital PCR Systems
| العنوان: | Poisson Plus Quantification for Digital PCR Systems |
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| المؤلفون: | Swapnonil Banerjee, Michael Pallas, Patricia Hegerich, Thomas Wessel, Nivedita Sumi Majumdar |
| المصدر: | Scientific Reports Scientific Reports, Vol 7, Iss 1, Pp 1-10 (2017) |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, Multidisciplinary, lcsh:R, 010401 analytical chemistry, Volume variation, Monte Carlo method, lcsh:Medicine, Empirical measure, Poisson distribution, Chip, 01 natural sciences, Sizing, Article, 0104 chemical sciences, 03 medical and health sciences, symbols.namesake, 030104 developmental biology, symbols, Partition (number theory), lcsh:Q, Digital polymerase chain reaction, lcsh:Science, Biological system, Mathematics |
| الوصف: | Digital PCR, a state-of-the-art nucleic acid quantification technique, works by spreading the target material across a large number of partitions. The average number of molecules per partition is estimated using Poisson statistics, and then converted into concentration by dividing by partition volume. In this standard approach, identical partition sizing is assumed. Violations of this assumption result in underestimation of target quantity, when using Poisson modeling, especially at higher concentrations. The Poisson-Plus Model accommodates for this underestimation, if statistics of the volume variation are well characterized. The volume variation was measured on the chip array based QuantStudio 3D Digital PCR System using the ROX fluorescence level as a proxy for effective load volume per through-hole. Monte Carlo simulations demonstrate the efficacy of the proposed correction. Empirical measurement of model parameters characterizing the effective load volume on QuantStudio 3D Digital PCR chips is presented. The model was used to analyze digital PCR experiments and showed improved accuracy in quantification. At the higher concentrations, the modeling must take effective fill volume variation into account to produce accurate estimates. The extent of the difference from the standard to the new modeling is positively correlated to the extent of fill volume variation in the effective load of your reactions. |
| تدمد: | 2045-2322 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::459bf1656f609dc513ed0ce657607d07 https://pubmed.ncbi.nlm.nih.gov/28851915 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....459bf1656f609dc513ed0ce657607d07 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20452322 |
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