ArticleNature protocols2014
Quadratic isothermal amplification for the detection of microRNA.
Article in Nature protocols, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 61 citations in OpenAlex.
- Inferring single-cell and spatial microRNA activity from transcriptomics data.Communications biology · 2025Article
- Ultrasensitive Detection of MicroRNA in Human Saliva via Rolling Circle Amplification Using a DNA-Decorated Graphene Oxide Sensor.ACS omega · 2023Article
- Isothermal exponential amplification reactions triggered by circular templates (cEXPAR) targeting miRNA.Molecular biology reports · 2023Article
- CRISPR-Cas12a based internal negative control for nonspecific products of exponential rolling circle amplification.Nucleic acids research · 2020Article
- Label-free detection of microRNA: two-stage signal enhancement with hairpin assisted cascade isothermal amplification and light-up DNA-silver nanoclusters.Mikrochimica acta · 2020Article
- Detection of Several Homologous MicroRNAs by a Single Smart Probe System Consisting of Linear Nucleic Acid Blockers.Molecules (Basel, Switzerland) · 2019Article
- Sensitively distinguishing intracellular precursor and mature microRNA abundance.Chemical science · 2019Article
- Microfluidic System for Detection of Viral RNA in Blood Using a Barcode Fluorescence Reporter and a Photocleavable Capture Probe.Analytical chemistry · 2017Article
- Multiplex quantitative analysis of microRNA expression via exponential isothermal amplification and conformation-sensitive DNA separation.Scientific reports · 2017Article
- MnOACS applied materials & interfaces · 2017Article
- Label-Free Isothermal Amplification Assay for Specific and Highly Sensitive Colorimetric miRNA Detection.ACS omega · 2016Article
- A conformation-induced fluorescence method for microRNA detection.Nucleic acids research · 2016Article
Corrections and comments
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Authors and funding
9 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
This protocol describes an isothermal amplification approach for ultrasensitive detection of specific microRNAs (miRNAs). It achieves this level of sensitivity through quadratic amplification of the target oligonucleotide by using a Bst DNA polymerase-induced strand-displacement reaction and a lambda exonuclease-aided recycling reaction. First, the target miRNA binds to a specifically designed molecular beacon, causing it to become a fluorescence emitter. A primer then binds to the activated beacon, and Bst polymerase initiates the synthesis of a double-stranded DNA segment templated on the molecular beacon. This causes the concomitant release of the target miRNA from the beacon--the first round of 'recycling'. Second, the duplex beacon thus produced is a suitable substrate for a nicking enzyme present in solution. After the duplex beacon is nicked, the lambda exonuclease digests the beacon and releases the DNA single strand just synthesized, which is complementary to the molecular beacon, inducing the second round of recycling. The miRNA detection limit of this protocol is 10 fmol at 37 °C and 1 amol at 4 °C. This approach also affords high selectivity when applied to miRNA extracted from MCF-7 and PC3 cell lines and even from breast cancer tissue samples. Upon isolation of miRNA, the detection process can be completed in ∼2 h.
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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.