{"doi":"10.1093/nar/29.9.e45","title":"A new mathematical model for relative quantification in real-time RT-PCR","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.","journal":"Nucleic Acids Research","year":2001,"id":9469,"datarank":26.283780843651364,"base_score":10.453543504793787,"endowment":10.453543504793787,"self_citation_contribution":1.5680315257190682,"citation_network_contribution":24.715749317932296,"self_endowment_contribution":1.5680315257190682,"citer_contribution":24.715749317932296,"corpus_percentile":99.5,"corpus_rank":129,"citation_count":34666,"citer_count":192,"citers_with_citation_signal":192,"citers_with_endowment":192,"datacite_reuse_total":0,"is_dataset":false,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2001-05-01","authors":[{"id":29251,"name":"Michael W. Pfaffl","orcid":"0000-0002-3192-1019","position":1,"is_corresponding":false}],"reference_count":26,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-03-01T18:20:47.508186Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}