biostudies-literatureOliveira-Jr GPNCATS NIH HHSNHLBI NIH HHSNCI NIH HHSNational Institutes of HealthNational Institute of General Medical SciencesNIGMS NIH HHS113307https://www.ebi.ac.uk/biostudies/studies/S-EPMC8461749biostudies-literatureUnknown189MicroRNAs are short, non-coding RNA sequences involved in gene expression regulation. Quantification of miRNAs in biological fluids involves time consuming and laborious methods such as Northern blotting or PCR-based techniques. Molecular beacons (MB) are an attractive means for rapid detection of miRNAs, although the need for sophisticated readout methods limits their use in research and clinical settings. Here, we introduce a novel method based on delayed electrophoretic mobility, as a quantitative means for detection of miRNAs-MB hybridization. Upon hybridization with the target miRNAs, MB form a fluorescent duplex with reduced electrophoretic mobility, thus bypassing the need for additional staining. In addition to emission of light, the location of the fluorescent band on the gel actsBiosensors & bioelectronicsElectrophoretic mobility shift as a molecular beacon-based readout for miRNA detection.PMC8461749UG3HL147353R01 CA218500UG3 TR002881R35 GM119537UG3TR002881RO1CA218500UG3 HL147353UH3 TR002881Tigges JHansen CHPinckney BJones JMurphy-Thornley MGhiran ICThompson LBarbosa RHLu SOliveira-Jr GPWong WPfalseElectrophoretic mobility shift as a molecular beacon-based readout for miRNA detection.MicroRNAs are short, non-coding RNA sequences involved in gene expression regulation. Quantification of miRNAs in biological fluids involves time consuming and laborious methods such as Northern blotting or PCR-based techniques. Molecular beacons (MB) are an attractive means for rapid detection of miRNAs, although the need for sophisticated readout methods limits their use in research and clinical settings. Here, we introduce a novel method based on delayed electrophoretic mobility, as a quantitative means for detection of miRNAs-MB hybridization. Upon hybridization with the target miRNAs, MB form a fluorescent duplex with reduced electrophoretic mobility, thus bypassing the need for additional staining. In addition to emission of light, the location of the fluorescent band on the gel acts2021-01-01T00:00:00Z2021 Oct2025-04-04T11:55:53.294Z2025-04-04T11:55:53.294ZS-EPMC84617493406233410.1016/j.bios.2021.113307