ReviewJournal of biomolecular techniques : JBT2004
Pitfalls of quantitative real-time reverse-transcription polymerase chain reaction.
Review in Journal of biomolecular techniques : JBT, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 412 papers, 2 of them syntheses that pooled it.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
412 citing papers in PubMed, 2 syntheses or guidelines pooled it.
- Systematic review on the association between respiratory virus real-time PCR cycle threshold values and clinical presentation or outcomes.The Journal of antimicrobial chemotherapy · 2021Pooled it
- Using Cell Cultures for the Investigation of Treatments for Attention Deficit Hyperactivity Disorder: A Systematic Review.Current neuropharmacology · 2019Pooled it
- Identification of Reference Genes and Their Application in Analysis of Ginsenoside Biosynthesis Genes in American Ginseng Under Abiotic Stresses and Hormone Treatments.Plants (Basel, Switzerland) · 2026Article
- Computational Method Using Attribute-Aware Message Passing and Graph Convolutional Network for Potential miRNA-Disease Association Prediction.International journal of molecular sciences · 2026Article
- Development of a new TaqMan real-time PCR for detecting Psittacine adenovirus 1 (PsAdV-1).BMC veterinary research · 2026Article
- Genome-wide identification, evolution, and expression pattern analysis of the LEA gene family in Tartary Buckwheat (Fagopyrum tataricum).BMC genomics · 2026Article
- Modern RNA Quantification Methods: From RT-qPCR to Advanced Microscopy.The journal of physical chemistry. B · 2026Review
- Mitochondrial miRNA- miR-181c-5p and mitomiR-106a-5p levels as indicators in cardiovascular disease patients.Molecular biology reports · 2026Article
- Reporting quality of quantitative polymerase chain reaction (qPCR) methods in scientific publications.Research integrity and peer review · 2026Article
- Screening and Stability Validation of RT-qPCR Reference Genes inInternational journal of molecular sciences · 2026Article
- Reference Gene Stability inInternational journal of molecular sciences · 2026Article
- Transcriptome-Based Identification of Reference Genes for Expression Analysis in Cassava UnderPlants (Basel, Switzerland) · 2025Article
- Dual-Emission FRET-PCR Outperforms SYBR Green and EvaGreen for Accurate Discrimination of Primary Canine Dermatophytes:Journal of fungi (Basel, Switzerland) · 2025Article
- Establishment of Singleplex and Duplex TaqMan RT-qPCR Detection Systems for Strawberry Mottle Virus (SMoV) and Strawberry Vein Banding Virus (SVBV).Plants (Basel, Switzerland) · 2025Article
- Article
- Machine learning selection of basement membrane-associated genes and development of a predictive model for kidney fibrosis.Scientific reports · 2025Article
- Reducing dextran sodium sulfate interference with gene expression quantification in a mouse model of colitis.Journal of animal science · 2025Article
- Cost-Effective and Sensitive Probe-Based Universal (PBU) qPCR for MicroRNA Quantification in Plants.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Evaluation of candidate reference genes for gene expression research inFrontiers in insect science · 2025Article
- Retrospective detection of severe fever with thrombocytopenia syndrome virus (SFTSV) revises timeline of emergence in Thailand.Virus evolution · 2025Article
352 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Polymerase chain reaction (PCR)-based assays can target either DNA (the genome) or RNA (the transcriptome). Targeting the genome generates robust data that are informative and, most importantly, generally applicable. This is because the information contained within the genome is context-independent; i.e., generally, every normal cell contains the same DNA sequence--the same mutations and polymorphisms. The transcriptome, on the other hand, is context-dependent; i.e., the mRNA complement and level varies with physiology, pathology, or development. This makes the information contained within the transcriptome intrinsically flexible and variable. If this variability is combined with the technical limitations inherent in any reverse-transcription (RT)-PCR assay, it can be difficult to achieve not just a technically accurate but a biologically relevant result. Template quality, operator variability, the RT step itself, and subjectivity in data analysis and reporting are just a few technical aspects that make real-time RT-PCR appear to be a fragile assay that makes accurate data interpretation difficult. There can be little doubt that in the future, transcriptome-based analysis will become a routine technique. However, for the time being it remains a research tool, and it is important to recognize the considerable pitfalls associated with transcriptome analysis, with the successful application of RTPCR depending on careful experimental design, application, and validation.
Indexed as
Identifiers
15331581PMC2291693What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.