Multiple stem-loop primers induced cascaded loop-mediated isothermal amplification for direct recognition and specific detection of circular RNAs
Xin Liu, Pengbo Zhang, Kejian Gao, Xiaoyu Wang, Zhengping Li
Abstract
DNA polymerase, the rationally designed SLPs can directly recognize the target circRNA and generate a large amount of double stem-loop DNAs which can initiate a cascaded LAMP reaction with improved efficiency. The use of additional SLPs can accelerate the reaction rate; as a result, the LAMP reaction can be finished within 35 min which is much quicker than traditional LAMP. The proposed method can detect as low as 1 fM circRNA and has been successfully applied to the detection of circRNAs in total RNA samples extracted from cell lines.
Topics & Concepts
Loop-mediated isothermal amplificationCircular RNAmicroRNAStem-loopRNAComputational biologyBiologyDNAMultiple displacement amplificationLong non-coding RNAPolymerase chain reactionCell biologyMolecular biologyGeneGeneticsDNA extractionCircular RNAs in diseasesMicroRNA in disease regulationExtracellular vesicles in disease