ArticlePloS one2014
3' IsomiR species and DNA contamination influence reliable quantification of microRNAs by stem-loop quantitative PCR.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed.
- Genome-wide microRNA analysis of HPV-positive self-samples yields novel triage markers for early detection of cervical cancer.International journal of cancer · 2019Trial
- Article
- 3'IsomiR Species Composition Affects Reliable Quantification of miRNA/isomiR Variants by Poly(A) RT-qPCR: Impact on Small RNA-Seq Profiling Validation.International journal of molecular sciences · 2023Article
- One locus, several functional RNAs-emerging roles of the mechanisms responsible for the sequence variability of microRNAs.Biologia futura · 2023Review
- MiR92b-3p synthetic analogue impairs zebrafish embryonic development, leading to ocular defects, decreased movement and hatching rate, and increased mortality.Journal of applied genetics · 2023Article
- The Role of MicroRNAs in Liver Functioning: from Biogenesis to Therapeutic Approaches (Review).Sovremennye tekhnologii v meditsine · 2023Review
- Rapid and accurate quantification of isomiRs by RT-qPCR.Scientific reports · 2022Article
- miRNAs and isomiRs: Serum-Based Biomarkers for the Development of Intellectual Disability and Autism Spectrum Disorder in Tuberous Sclerosis Complex.Biomedicines · 2022Article
- Implication of genetic variants in primary microRNA processing sites in the risk of multiple sclerosis.EBioMedicine · 2022Article
- Quantification of microRNA editing using two-tailed RT-qPCR for improved biomarker discovery.RNA (New York, N.Y.) · 2021Article
- Improved protocol for plasma microRNA extraction and comparison of commercial kits.Biochemia medica · 2021Article
- Review
- Accurate Detection of Target MicroRNA in Mixed Species of High Sequence Homology Using Target-Protection Rolling Circle Amplification.ACS omega · 2021Article
- Effective tools for RNA-derived therapeutics: siRNA interference or miRNA mimicry.Theranostics · 2021Review
- The MicroRNA Family Gets Wider: The IsomiRs Classification and Role.Frontiers in cell and developmental biology · 2021Review
- Quantification of purified endogenous miRNAs with high sensitivity and specificity.Nature communications · 2020Article
- On the expanding roles of tRNA fragments in modulating cell behavior.Nucleic acids research · 2020Review
- Stumbling on elusive cargo: how isomiRs challenge microRNA detection and quantification, the case of extracellular vesicles.Journal of extracellular vesicles · 2020Article
- Article
- A Comprehensive Approach to Sequence-oriented IsomiR annotation (CASMIR): demonstration with IsomiR profiling in colorectal neoplasia.BMC genomics · 2018Article
Corrections and comments
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
MicroRNAs (miRNAs) are ∼20-24 nucleotide-long regulatory RNAs that have been proven to play important roles in many cellular processes. Since their discovery, a number of different techniques have been developed to detect and accurately quantify them. For individual mature miRNA measurements, quantitative stem-loop real-time PCR represents a widely used method. Although there are some data on optimization of this technique, there are still many factors that have not been investigated yet. In this study, we have thoroughly optimized this technique and pointed out several important factors that influence reliable quantification. First, we found that total RNA input can affect the measurements. Second, our data showed that carryover DNA contamination could also mislead the detection in a sequence-specific manner. Additionally, we provided evidence that different 3' isomiR species of a particular miRNA can be reverse transcribed and cross-detected even by specifically targeted assays. Besides these, we have investigated the measurement of reaction efficiencies from total RNA samples and the accuracy of simultaneous reverse transcription reactions for increasing reliability and cost effectiveness without the loss of sensitivity and specificity. In summary, we provide a detailed, refined protocol for reliable detection of microRNA species by quantitative stem-loop PCR.
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