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

STAR Protoc

Zhejiang Provincial Key Laboratory of Precision Diagnosis and Therapy for Major Gynecological Diseases, Women's Hospital and Institute of Translational Medicine, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, China. Electronic address:

Published: December 2024

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. and Sun et al..

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11574804PMC
http://dx.doi.org/10.1016/j.xpro.2024.103435DOI Listing

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