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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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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author Mengqian Yu
Pengyuan Liu
Yan Lu
author_facet Mengqian Yu
Pengyuan Liu
Yan Lu
author_sort Mengqian Yu
collection DOAJ
description 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.
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series STAR Protocols
spelling doaj-art-6214b8314b3c40a59cf5fcc376527de02024-11-20T05:08:12ZengElsevierSTAR Protocols2666-16672024-12-0154103435Protocol for detection of tRNA-derived fragments in cells, tissues, and plasmaMengqian Yu0Pengyuan Liu1Yan Lu2Zhejiang 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; Corresponding authorDepartment of Respiratory Medicine, Sir Run Run Shaw Hospital and Institute of Translational Medicine, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310016, ChinaZhejiang 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; Corresponding authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2666166724006002CancerMolecular BiologyGene Expression
spellingShingle Mengqian Yu
Pengyuan Liu
Yan Lu
Protocol for detection of tRNA-derived fragments in cells, tissues, and plasma
STAR Protocols
Cancer
Molecular Biology
Gene Expression
title Protocol for detection of tRNA-derived fragments in cells, tissues, and plasma
title_full Protocol for detection of tRNA-derived fragments in cells, tissues, and plasma
title_fullStr Protocol for detection of tRNA-derived fragments in cells, tissues, and plasma
title_full_unstemmed Protocol for detection of tRNA-derived fragments in cells, tissues, and plasma
title_short Protocol for detection of tRNA-derived fragments in cells, tissues, and plasma
title_sort protocol for detection of trna derived fragments in cells tissues and plasma
topic Cancer
Molecular Biology
Gene Expression
url http://www.sciencedirect.com/science/article/pii/S2666166724006002
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