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Biol."],"published-print":{"date-parts":[[2025]]},"abstract":"<jats:p>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 \u03bcg 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\u00adty suitable for subsequent RNA NGS applications.<\/jats:p>","DOI":"10.14712\/fb2025071020088","type":"journal-article","created":{"date-parts":[[2025,7,8]],"date-time":"2025-07-08T16:24:03Z","timestamp":1751991843000},"page":"88-94","source":"Crossref","is-referenced-by-count":2,"title":["Semi-automated RNA Isolation from Tempus Blood RNA Tubes Using the Magcore Plus II Instrument"],"prefix":"10.14712","volume":"71","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4205-2020","authenticated-orcid":false,"given":"Marta","family":"\u010cern\u00e1","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3463-8320","authenticated-orcid":false,"given":"Barbora","family":"\u0160\u0165astn\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pavel","family":"Pe\u0161ek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-7205-7287","authenticated-orcid":false,"given":"Ta\u0165\u00e1na","family":"Pt\u00e1\u010dkov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2816-6769","authenticated-orcid":false,"given":"Mark\u00e9ta","family":"Janatov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2413-1542","authenticated-orcid":false,"given":"Jana","family":"Soukupov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7390-4751","authenticated-orcid":false,"given":"Kl\u00e1ra","family":"Hor\u00e1\u010dkov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4806-9854","authenticated-orcid":false,"given":"Petra","family":"Kleiblov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"5727","published-online":{"date-parts":[[2025,7,8]]},"reference":[{"key":"fb2025071020088_1","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1038\/nrg.2016.10","volume":"17","author":"Byron","year":"2016","journal-title":"Nat. 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