Standard operating procedure for next generation sequencing of RNA isolated form extracellular vesicles within the nanostructurome methods pipeline

Kovachevikj, Miona (avtor), Kralj-Iglič, Veronika (avtor), Jakše, Jernej (avtor), Jeseničnik, Taja (avtor)

The standard operating procedure (SOP) for next generation sequencing (NGS) of RNA from extracellular vesicles is presented. The SOP enables purification of small or total RNAs from isolated extracellular vesicles. Next, the SOP describes the sequencing of RNAs using NGS sequencing platform IonTorrent. In the presented protocol we define all the steps from isolation of RNA to sequencing data management, together with the cautions and checkpoints crucial for obtaining reliable high-quality sequencing reads, suitable for further analysis. NGS enables high-throughput DNA and RNA sequencing, offering accuracy, scalability, and cost-efficiency. RNA sequencing is widely used for analysing gene expression, alternative splicing, and non-coding RNAs. NGS also facilitates the study of extracellular vesicles (EVs), which play a key role in intercellular communication, especially in plant-pathogen interactions. Insights gained could inform novel disease management strategies, such as Spray-Induced Gene Silencing, for improved crop protection.

• Jezik: Angleški jezik
• Ključne besede: next generation sequencing, extracellular vesicles, RNA-seq, total RNA, small RNA, IonTorrent
• Vrsta gradiva: Znanstveno delo
• Tipologija: 1.08 - Objavljeni znanstveni prispevek na konferenci
• Organizacija: ZF - Zdravstvena fakulteta ; BF - Biotehniška fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2025
• Št. strani: Str. 18-24
• PID: 20.500.12556/RUL-169993
• UDK: 577
• DOI: 10.55295/PSL.12.2025.III3
• COBISS.SI-ID: 240973827
• Datum objave v RUL: 01.07.2025

Gradivo je del monografije

• Naslov: Socratic Lectures : 12th international symposium, Ljubljana, 11. 1. 2025
• Uredniki: Veronika Kralj-Iglič, Yelena Istileulova, Anna Romolo
• Kraj izida: Ljubljana
• Založnik: University of Ljubljana Press
• Leto izida: 2025
• ISBN: 978-961-297-597-5
• COBISS.SI-ID: 237747715

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Projekti

• Financer: ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
• Številka projekta: P4-0077-2019
• Naslov: Kmetijske rastline - genetika in sodobne tehnologije

• Financer: ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
• Številka projekta: 802-12/2024-5, J3-60063,
• Naslov: Nanostructurome