Semi-automated RNA Isolation from Tempus Blood RNA Tubes Using the Magcore Plus II Instrument

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00064165%3A_____%2F25%3A10499691" target="_blank" >RIV/00064165:_____/25:10499691 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11110/25:10499691 RIV/00216208:11310/25:10499691

Výsledek na webu

<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=t-.9wU.kBZ" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=t-.9wU.kBZ</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.14712/fb2025071020088" target="_blank" >10.14712/fb2025071020088</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Semi-automated RNA Isolation from Tempus Blood RNA Tubes Using the Magcore Plus II Instrument

Popis výsledku v původním jazyce

High-throughput, precise and cost-effective isolation of high-quality RNA is essential for the growing number of RNA-based next-generation sequencing (NGS) analyses. Manual RNA isolation provides sufficient quality but requires significant hands-on time and carries an increased risk of contamination and sample misidentification. Here we describe a semi-automated protocol for the isolation of high-quality total RNA from 3 ml of peripheral blood collected in Tempus Blood RNA Tubes. The isolation can be performed either from the total volume of 9 ml of Tempus blood lysate or from smaller volumes (6 and 3 ml, respectively) using the MagCore triXact RNA Kit on the MagCore Plus II automated nucleic acid extractor, which allows RNA isolation in single tubes. The original isolation protocol (#631) for whole blood RNA isolation was customized by the manufacturer (#631T) by omitting the cell lysis step. After optimizing the process, we compared the yield and quality of 760 RNA samples isolated manually or by semi-automated methods. We conclude that RNA isolation using the semi-automated MagCore protocol yields 5-10 μg of total RNA from 6 ml of lysate (2 ml of peripheral blood), which is almost comparable in quantity and quality to manual isolation. In addition, we show that the remaining 3 ml of lysate is sufficient for backup re-isolation. Our semi-automated RNA protocol reduces hands-on time without increasing costs and yields bulky total RNA of a quali-ty suitable for subsequent RNA NGS applications.

Název v anglickém jazyce

Semi-automated RNA Isolation from Tempus Blood RNA Tubes Using the Magcore Plus II Instrument

Popis výsledku anglicky

High-throughput, precise and cost-effective isolation of high-quality RNA is essential for the growing number of RNA-based next-generation sequencing (NGS) analyses. Manual RNA isolation provides sufficient quality but requires significant hands-on time and carries an increased risk of contamination and sample misidentification. Here we describe a semi-automated protocol for the isolation of high-quality total RNA from 3 ml of peripheral blood collected in Tempus Blood RNA Tubes. The isolation can be performed either from the total volume of 9 ml of Tempus blood lysate or from smaller volumes (6 and 3 ml, respectively) using the MagCore triXact RNA Kit on the MagCore Plus II automated nucleic acid extractor, which allows RNA isolation in single tubes. The original isolation protocol (#631) for whole blood RNA isolation was customized by the manufacturer (#631T) by omitting the cell lysis step. After optimizing the process, we compared the yield and quality of 760 RNA samples isolated manually or by semi-automated methods. We conclude that RNA isolation using the semi-automated MagCore protocol yields 5-10 μg of total RNA from 6 ml of lysate (2 ml of peripheral blood), which is almost comparable in quantity and quality to manual isolation. In addition, we show that the remaining 3 ml of lysate is sufficient for backup re-isolation. Our semi-automated RNA protocol reduces hands-on time without increasing costs and yields bulky total RNA of a quali-ty suitable for subsequent RNA NGS applications.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30100 - Basic medicine

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Folia Biologica

ISSN

0015-5500

e-ISSN

2533-7602

Svazek periodika

71

Číslo periodika v rámci svazku

2

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

7

Strana od-do

88-94

Kód UT WoS článku

001529657600005

EID výsledku v databázi Scopus

2-s2.0-105010975104