ArticleBMC medical genomics2013
A novel method for the normalization of microRNA RT-PCR data.
Article in BMC medical genomics, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers, 1 of them a synthesis that pooled it.
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Who cites it
32 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Bioinformatics analysis of the expression and role of microRNA-221-3p in head and neck squamous cell carcinoma.BMC cancer · 2021Pooled it
- Circulating miR-10b-5p as a candidate biomarker of atrial fibrillation recurrence after catheter ablation: a two-phase translational study.Europace : European pacing, arrhythmias, and cardiac electrophysiology : journal of the working groups on cardiac pacing, arrhythmias, and cardiac cellular electrophysiology of the European Society of Cardiology · 2026Article
- Under heavy metal stress: genomic characterization and expression profiles of miRNA families in Phaseolus vulgaris.BMC plant biology · 2026Article
- Genome-wide analysis of Phaseolus vulgaris L. miR172 gene family members under heavy metal stress.BMC genomics · 2026Article
- Evaluation of Suitable Reference Gene During the Development of Paired or Unpaired FemalePathogens (Basel, Switzerland) · 2025Article
- The effect of Entamoeba histolytica lectin antigen and microRNA-643 on the development of microsatellite instability (MSI) in colorectal adenocarcinoma.BMC cancer · 2025Article
- miR-1233-3p Inhibits Angiopoietin-1-Induced Endothelial Cell Survival, Migration, and Differentiation.Cells · 2025Article
- Association between Hypertension with and without Left Ventricular Hypertrophy and the Expression of a Panel of microRNAs in an Iranian Population.Research in heart yield and translational medicine · 2025Article
- Genome-wide analysis of miR172-mediated response to heavy metal stress in chickpea (Cicer arietinum L.): physiological, biochemical, and molecular insights.BMC plant biology · 2024Article
- Effect of green coffee on miR-133a, miR-155 and inflammatory biomarkers in obese individuals.Diabetology & metabolic syndrome · 2024Article
- InterOpt: Improved gene expression quantification in qPCR experiments using weighted aggregation of reference genes.iScience · 2023Article
- An exploratory approach to identify microRNAs as circulatory biomarker candidates for epilepsy-associated psychiatric comorbidities in an electrical post-status epilepticus model.Scientific reports · 2023Article
- Germline variants in tumor suppressor FBXW7 lead to impaired ubiquitination and a neurodevelopmental syndrome.American journal of human genetics · 2022Article
- Measurements Methods for the Development of MicroRNA-Based Tests for Cancer Diagnosis.International journal of molecular sciences · 2021Review
- Removal of COVID-19 Spike Protein, Whole Virus, Exosomes, and Exosomal MicroRNAs by the Hemopurifier® Lectin-Affinity Cartridge in Critically Ill Patients With COVID-19 Infection.Frontiers in medicine · 2021Article
- Analysis of Reciprocally Dysregulated miRNAs in Eutopic Endometrium Is a Promising Approach for Low Invasive Diagnostics of Adenomyosis.Diagnostics (Basel, Switzerland) · 2020Article
- Urine microRNA Profiling Displays miR-125a Dysregulation in Children with Fragile X Syndrome.Cells · 2020Article
- Conserved regulation of neurodevelopmental processes and behavior by FoxP in Drosophila.PloS one · 2019Article
- Identification of Endogenous Control miRNAs for RT-qPCR in T-Cell Acute Lymphoblastic Leukemia.International journal of molecular sciences · 2018Article
- The choice of endogenous controls in exosomal microRNA assessments from biofluids.Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine · 2016Review
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundMicroRNAs (miRNAs) are short non-coding RNA molecules that regulate mRNA transcript levels and translation. Deregulation of microRNAs is indicated in a number of diseases and microRNAs are seen as a promising target for biomarker identification and drug development. miRNA expression is commonly measured by microarray or real-time polymerase chain reaction (RT-PCR). The findings of RT-PCR data are highly dependent on the normalization techniques used during preprocessing of the Cycle Threshold readings from RT-PCR. Some of the commonly used endogenous controls themselves have been discovered to be differentially expressed in various conditions such as cancer, making them inappropriate internal controls.
methodsWe demonstrate that RT-PCR data contains a systematic bias resulting in large variations in the Cycle Threshold (CT) values of the low-abundant miRNA samples. We propose a new data normalization method that considers all available microRNAs as endogenous controls. A weighted normalization approach is utilized to allow contribution from all microRNAs, weighted by their empirical stability.
resultsThe systematic bias in RT-PCR data is illustrated on a microRNA dataset obtained from primary cutaneous melanocytic neoplasms. We show that through a single control parameter, this method is able to emulate other commonly used normalization methods and thus provides a more general approach. We explore the consistency of RT-PCR expression data with microarray expression by utilizing a dataset where both RT-PCR and microarray profiling data is available for the same miRNA samples.
conclusionsA weighted normalization method allows the contribution of all of the miRNAs, whether they are highly abundant or have low expression levels. Our findings further suggest that the normalization of a particular miRNA should rely on only miRNAs that have comparable expression levels.
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