{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T10:29:20Z","timestamp":1786444160942,"version":"build-2736575974"},"posted":{"date-parts":[[2026,4,7]]},"group-title":"Neuroscience","reference-count":20,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T00:00:00Z","timestamp":1775520000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health grant","doi-asserted-by":"crossref","award":["R03 AG078787"],"award-info":[{"award-number":["R03 AG078787"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health grant","doi-asserted-by":"crossref","award":["National Institutes of Health grant"],"award-info":[{"award-number":["National Institutes of Health grant"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health grant","doi-asserted-by":"crossref","award":["R01 AG086396"],"award-info":[{"award-number":["R01 AG086396"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,8,8]]},"abstract":"<jats:title>SUMMARY<\/jats:title>\n                <jats:p>Nuclei isolation from myelin-rich adult mouse brain regions remains challenging for single-nucleus RNA sequencing because myelin and debris can reduce nuclei quality. We describe an optimized protocol for mouse hippocampi and cerebella using tube-and-pestle homogenization, low-volume sucrose-gradient pelleting with a standard benchtop centrifuge, and magnetic enrichment as the recommended final cleanup step to reduce debris\/non-nuclear carryover. Under the tested conditions, the workflow produces intact, debris-reduced nuclei and supports downstream 10x Genomics Flex and PARSE WT library preparation.<\/jats:p>\n                <jats:sec>\n                  <jats:title>Graphical abstract<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" orientation=\"portrait\" fig-type=\"figure\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"716374v2_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Highlights<\/jats:title>\n                  <jats:list list-type=\"bullet\">\n                    <jats:list-item>\n                      <jats:p>Benchtop sucrose-gradient pelleting enables rapid nuclei purification from myelin-rich adult mouse brain<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>Scales across tissue inputs (e.g., hippocampus \u223c15\u201320 mg; cerebellum \u223c50\u201370 mg) without ultracentrifugation or 15 mL gradients<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>Magnetic enrichment as the recommended final cleanup step further reduces myelin\/debris carryover and is compatible with 10x Flex and PARSE WT workflows.<\/jats:p>\n                    <\/jats:list-item>\n                  <\/jats:list>\n                <\/jats:sec>","DOI":"10.64898\/2026.04.03.716374","type":"posted-content","created":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T19:25:20Z","timestamp":1775589920000},"source":"Crossref","is-referenced-by-count":0,"title":["Myelin-Free Nuclei Isolation from Mouse Hippocampus and Cerebellum for snRNA-Seq with Benchtop Gradient Centrifugation"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-0610-3380","authenticated-orcid":false,"given":"Benu","family":"George","sequence":"first","affiliation":[{"name":"Department of Neurology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Braedon Q","family":"Kirkpatrick","sequence":"additional","affiliation":[{"name":"Department of Neurology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4061-6278","authenticated-orcid":false,"given":"Victor","family":"Kilonzo","sequence":"additional","affiliation":[{"name":"Department of Neurology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4746-0392","authenticated-orcid":false,"given":"Qiang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Neurology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026081102350507000_2026.04.03.716374v2.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2022.101944"},{"key":"2026081102350507000_2026.04.03.716374v2.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2024.103395"},{"key":"2026081102350507000_2026.04.03.716374v2.3","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-020-00472-3"},{"key":"2026081102350507000_2026.04.03.716374v2.4","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4407"},{"key":"2026081102350507000_2026.04.03.716374v2.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.crmeth.2026.101337"},{"key":"2026081102350507000_2026.04.03.716374v2.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2021.100844"},{"key":"2026081102350507000_2026.04.03.716374v2.7","doi-asserted-by":"publisher","DOI":"10.3389\/fncir.2020.611841"},{"key":"2026081102350507000_2026.04.03.716374v2.8","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2014.00450"},{"key":"2026081102350507000_2026.04.03.716374v2.9","doi-asserted-by":"publisher","DOI":"10.1126\/science.aam8999"},{"key":"2026081102350507000_2026.04.03.716374v2.10","unstructured":"10x Genomics. 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