A new mathematical model for relative quantification in real-time RT-PCR. Export

@article{citeulike:278954, abstract = {Use of the real-time polymerase chain reaction (PCR) to amplify cDNA products reverse transcribed from mRNA is on the way to becoming a routine tool in molecular biology to study low abundance gene expression. Real-time PCR is easy to perform, provides the necessary accuracy and produces reliable as well as rapid quantification results. But accurate quantification of nucleic acids requires a reproducible methodology and an adequate mathematical model for data analysis. This study enters into the particular topics of the relative quantification in real-time RT-PCR of a target gene transcript in comparison to a reference gene transcript. Therefore, a new mathematical model is presented. The relative expression ratio is calculated only from the real-time PCR efficiencies and the crossing point deviation of an unknown sample versus a control. This model needs no calibration curve. Control levels were included in the model to standardise each reaction run with respect to RNA integrity, sample loading and inter-PCR variations. High accuracy and reproducibility (<2.5\% variation) were reached in LightCycler PCR using the established mathematical model.}, address = {Institute of Physiology, FML-Weihenstephan, Center of Life and Food Sciences, Technical University of Munich, Germany. pfaffl@weihenstephan.de}, author = {Pfaffl, M. W.}, citeulike-article-id = {278954}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/11328886}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=11328886}, day = {1}, issn = {1362-4962}, journal = {Nucleic acids research}, keywords = {pcr, quantify}, month = {May}, number = {9}, posted-at = {2009-02-02 17:54:12}, priority = {2}, title = {A new mathematical model for relative quantification in real-time RT-PCR.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/11328886}, volume = {29}, year = {2001} }

TY - JOUR ID - citeulike:278954 L3 - citeulike-article-id:278954 N2 - Use of the real-time polymerase chain reaction (PCR) to amplify cDNA products reverse transcribed from mRNA is on the way to becoming a routine tool in molecular biology to study low abundance gene expression. Real-time PCR is easy to perform, provides the necessary accuracy and produces reliable as well as rapid quantification results. But accurate quantification of nucleic acids requires a reproducible methodology and an adequate mathematical model for data analysis. This study enters into the particular topics of the relative quantification in real-time RT-PCR of a target gene transcript in comparison to a reference gene transcript. Therefore, a new mathematical model is presented. The relative expression ratio is calculated only from the real-time PCR efficiencies and the crossing point deviation of an unknown sample versus a control. This model needs no calibration curve. Control levels were included in the model to standardise each reaction run with respect to RNA integrity, sample loading and inter-PCR variations. High accuracy and reproducibility (<2.5% variation) were reached in LightCycler PCR using the established mathematical model. IS - 9 JF - Nucleic acids research SN - 1362-4962 AD - Institute of Physiology, FML-Weihenstephan, Center of Life and Food Sciences, Technical University of Munich, Germany. pfaffl@weihenstephan.de TI - A new mathematical model for relative quantification in real-time RT-PCR. VL - 29 KW - pcr KW - quantify AU - Pfaffl, MW PY - 2001/05/01/ UR - http://view.ncbi.nlm.nih.gov/pubmed/11328886 ER -