Protocol for detection of tRNA-derived fragments in cells, tissues, and plasma

Summary: tRNA-derived fragments (tRFs) are frequently dysregulated in cancers, and approaches for the detection of tRFs within biological samples are vital for their expression analysis and functional exploration. Here, we present a protocol for detecting tRFs using a modified TaqMan quantitative re...

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Bibliographic Details
Main Authors: Mengqian Yu, Pengyuan Liu, Yan Lu
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724006002
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Summary:Summary: tRNA-derived fragments (tRFs) are frequently dysregulated in cancers, and approaches for the detection of tRFs within biological samples are vital for their expression analysis and functional exploration. Here, we present a protocol for detecting tRFs using a modified TaqMan quantitative real-time PCR (qRT-PCR)-based technique, Dumbbell-PCR (Db-PCR). We describe steps for primer and adapter design, adapter-RNA ligation, and RNA detection. This protocol streamlines and enhances the precision of tRF quantification in cells, tissues, and plasma, facilitating a time-efficient and reliable assessment of their presence.For complete details on the use and execution of this protocol, please refer to Yu et al.1 and Sun et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667