ArticleRNA (New York, N.Y.)2013
A universal TaqMan-based RT-PCR protocol for cost-efficient detection of small noncoding RNA.
Article in RNA (New York, N.Y.), 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- A selective enrichment and specific probe terminal mediated strategy for highly sensitive detection of microRNAs.Nature communications · 2026Article
- The beauty of symmetry: siRNA phosphorodithioate modifications reduce stereocomplexity, ease analysis, and can improveMolecular therapy. Nucleic acids · 2024Article
- Inhibition of miR-200b-3p confers broad-spectrum resistance to viral infection by targeting TBK1.mBio · 2023Article
- OUHP: an optimized universal hairpin primer system for cost-effective and high-throughput RT-qPCR-based quantification of microRNA (miRNA) expression.Nucleic acids research · 2022Article
- Positive control synthesis method for COVID-19 diagnosis by one-step real-time RT-PCR.Clinica chimica acta; international journal of clinical chemistry · 2020Article
- Article
- Differential Expression of Maize and Teosinte microRNAs under Submergence, Drought, and Alternated Stress.Plants (Basel, Switzerland) · 2020Article
- Research progress in molecular biology related quantitative methods of MicroRNA.American journal of translational research · 2020Review
- RNA Structure Design Improves Activity and Specificity of trans-Splicing-Triggered Cell Death in a Suicide Gene Therapy Approach.Molecular therapy. Nucleic acids · 2018Article
- Multiplex quantitative analysis of microRNA expression via exponential isothermal amplification and conformation-sensitive DNA separation.Scientific reports · 2017Article
- Advanced Design of Dumbbell-shaped Genetic Minimal Vectors Improves Non-coding and Coding RNA Expression.Molecular therapy : the journal of the American Society of Gene Therapy · 2016Article
- Efficient production of superior dumbbell-shaped DNA minimal vectors for small hairpin RNA expression.Nucleic acids research · 2015Article
- Assessment of small RNA sorting into different extracellular fractions revealed by high-throughput sequencing of breast cell lines.Nucleic acids research · 2015Article
- Micro-RNAs in regenerating lungs: an integrative systems biology analysis of murine influenza pneumonia.BMC genomics · 2014Article
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Authors and funding
4 authors.
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Abstract
Several methods for the detection of RNA have been developed over time. For small RNA detection, a stem-loop reverse primer-based protocol relying on TaqMan RT-PCR has been described. This protocol requires an individual specific TaqMan probe for each target RNA and, hence, is highly cost-intensive for experiments with small sample sizes or large numbers of different samples. We describe a universal TaqMan-based probe protocol which can be used to detect any target sequence and demonstrate its applicability for the detection of endogenous as well as artificial eukaryotic and bacterial small RNAs. While the specific and the universal probe-based protocol showed the same sensitivity, the absolute sensitivity of detection was found to be more than 100-fold lower for both than previously reported. In subsequent experiments, we found previously unknown limitations intrinsic to the method affecting its feasibility in determination of mature template RISC incorporation as well as in multiplexing. Both protocols were equally specific in discriminating between correct and incorrect small RNA targets or between mature miRNA and its unprocessed RNA precursor, indicating the stem-loop RT-primer, but not the TaqMan probe, triggers target specificity. The presented universal TaqMan-based RT-PCR protocol represents a cost-efficient method for the detection of small RNAs.
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