Miniatura

Fitxers

835321.pdf (1.83 MB)

Tipus de document

Article

Versió

Versió publicada

Data de publicació

2023-07-29

Llicència de publicació

cc-by (c) Llorens-Revull M et al., 2023
Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/207048

Comparison of Extracellular Vesicle Isolation Methods for miRNA Sequencing

Títol de la revista

Autors

Director/Tutor

ISSN de la revista

Títol del volum

Recurs relacionat

https://doi.org/10.3390/ijms241512183

Resum

MicroRNAs (miRNAs) encapsulated in extracellular vesicles (EVs) are potential diagnostic and prognostic biomarkers. However, discrepancies in miRNA patterns and their validation are still frequent due to differences in sample origin, EV isolation, and miRNA sequencing methods. The aim of the present study is to find a reliable EV isolation method for miRNA sequencing, adequate for clinical application. To this aim, two comparative studies were performed in parallel with the same human plasma sample: (i) isolation and characterization of EVs obtained using three procedures: size exclusion chromatography (SEC), iodixanol gradient (GRAD), and its combination (SEC+GRAD) and (ii) evaluation of the yield of miRNA sequences obtained using NextSeq 500 (Illumina) and three miRNA library preparation protocols: NEBNext, NEXTFlex, and SMARTer smRNA-seq. The conclusion of comparison (i) is that recovery of the largest amount of EVs and reproducibility were attained with SEC, but GRAD and SEC+GRAD yielded purer EV preparations. The conclusion of (ii) is that the NEBNext library showed the highest reproducibility in the number of miRNAs recovered and the highest diversity of miRNAs. These results render the combination of GRAD EV isolation and NEBNext library preparation for miRNA retrieval as adequate for clinical applications using plasma samples.

Matèries

RNA, Marcadors bioquímics

Matèries (anglès)

RNA, Biochemical markers

Citació

Col·leccions

Articles publicats en revistes (Genètica, Microbiologia i Estadística)

Citació

LLORENS-REVULL, Meritxell, MARTÍNEZ-GONZÁLEZ, Brenda, QUER, Josep, ESTEBAN, Juan ignacio, NÚÑEZ-MORENO, Gonzalo, MÍNGUEZ, Pablo, BURGUI, Idoia, RAMOS-RUÍZ, Ricardo, SORIA, María eugenia, RICO, Angie, RIVEIRO-BARCIELA, Mar, SAULEDA, Silvia, PIRON, Maria, CORRALES, Irene, BORRÀS I SERRES, Francesc enric, RODRÍGUEZ-FRÍAS, Francisco, RANDO, Ariadna, RAMÍREZ-SERRA, Clara, CAMÓS, Silvia, DOMINGO, Esteban, BES, Marta, PERALES, Celia, COSTAFREDA SALVANY, M. isabel (maria isabel). Comparison of Extracellular Vesicle Isolation Methods for miRNA Sequencing. _International Journal of Molecular Sciences_. 2023. Vol. 24, núm. 15, pàgs. 1-15. [consulta: 21 de gener de 2026]. ISSN: 1661-6596. [Disponible a: https://hdl.handle.net/2445/207048]

Exportar metadades

JSON - METS

Compartir registre