{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T07:20:58Z","timestamp":1778224858547,"version":"3.51.4"},"reference-count":28,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2021,4,22]],"date-time":"2021-04-22T00:00:00Z","timestamp":1619049600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["ROI DK67536; RO1 DK103215"],"award-info":[{"award-number":["ROI DK67536; RO1 DK103215"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Societ\u00e0 Italiana Diabetologia (SID)","award":["FODIRI-MSD 2014\/2015"],"award-info":[{"award-number":["FODIRI-MSD 2014\/2015"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>The isolation of high-quality RNA from endocrine pancreas sections represents a considerable challenge largely due to the high ribonuclease levels. Laser capture microdissection (LCM) of mammalian islets, in association with RNA extraction protocols, has emerged as a feasible approach to characterizing their genetic and proteomic profiles. However, a validated protocol to obtain high-quality RNA from LCM-derived human pancreas specimens that is appropriate for next-generation sequencing analysis is still lacking. In this study, we applied four methods (Picopure extraction kit, Qiazol protocol, Qiazol + Clean-up kit, and RNeasy Microkit + Carrier) to extract RNA from human islets obtained from both non-diabetic individuals and patients with type 2 diabetes who had undergone partial pancreatectomy, as well as handpicked islets from both non-diabetic and diabetic organ donors. The yield and purity of total RNA were determined by 260\/280 absorbance using Nanodrop 100 and the RNA integrity number with a bioanalyzer. The results indicated that among the four methods, the RNeasy MicroKit + Carrier (Qiagen) provides the highest yield and purity.<\/jats:p>","DOI":"10.3390\/biom11050625","type":"journal-article","created":{"date-parts":[[2021,4,22]],"date-time":"2021-04-22T13:59:14Z","timestamp":1619099954000},"page":"625","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A Systematic Comparison of Protocols for Recovery of High-Quality RNA from Human Islets Extracted by Laser Capture Microdissection"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8665-8406","authenticated-orcid":false,"given":"Chiara M. A.","family":"Cefalo","sequence":"first","affiliation":[{"name":"Islet Cell Biology &amp; Regenerative Medicine, Joslin Diabetes Center, Department of Medicine, Brigham and Women\u2019s Hospital, Harvard Medical School, Harvard Stem Cell Institute, Boston, MA 02215, USA"},{"name":"Dipartimento di Scienze Mediche e Chirurgiche, Centro per le Malattie Endocrino-Metaboliche, Fondazione Policlinico Universitario Agostino Gemelli IRCCS, 00168 Rome, Italy"},{"name":"Dipartimento di Medicina e Chirurgia Traslazionale, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Teresa","family":"Mezza","sequence":"additional","affiliation":[{"name":"Islet Cell Biology &amp; Regenerative Medicine, Joslin Diabetes Center, Department of Medicine, Brigham and Women\u2019s Hospital, Harvard Medical School, Harvard Stem Cell Institute, Boston, MA 02215, USA"},{"name":"Dipartimento di Scienze Mediche e Chirurgiche, Centro per le Malattie Endocrino-Metaboliche, Fondazione Policlinico Universitario Agostino Gemelli IRCCS, 00168 Rome, Italy"},{"name":"Dipartimento di Medicina e Chirurgia Traslazionale, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7462-7792","authenticated-orcid":false,"given":"Andrea","family":"Giaccari","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Mediche e Chirurgiche, Centro per le Malattie Endocrino-Metaboliche, Fondazione Policlinico Universitario Agostino Gemelli IRCCS, 00168 Rome, Italy"},{"name":"Dipartimento di Medicina e Chirurgia Traslazionale, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5029-6119","authenticated-orcid":false,"given":"Rohit N.","family":"Kulkarni","sequence":"additional","affiliation":[{"name":"Islet Cell Biology &amp; Regenerative Medicine, Joslin Diabetes Center, Department of Medicine, Brigham and Women\u2019s Hospital, Harvard Medical School, Harvard Stem Cell Institute, Boston, MA 02215, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"998","DOI":"10.1126\/science.274.5289.998","article-title":"Laser capture microdissection","volume":"274","author":"Bonner","year":"1996","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.jprot.2016.09.002","article-title":"Proteomic profiling of human islets collected from frozen pancreata using laser capture microdissection","volume":"150","author":"Zhang","year":"2017","journal-title":"J. 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