{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T22:35:04Z","timestamp":1787956504112,"version":"build-2784847793"},"reference-count":24,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2018,6,19]],"date-time":"2018-06-19T00:00:00Z","timestamp":1529366400000},"content-version":"vor","delay-in-days":18,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100003202","name":"North American Spine Society","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100003202","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002732","name":"AOSpine","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002732","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["JOR Spine"],"published-print":{"date-parts":[[2018,6]]},"abstract":"<jats:p>The isolation of high\u2010quality RNA from the intervertebral disc and especially from the nucleus pulposus is challenging due to the low cellularity and high proteoglycan content of this tissue. In this study, we report a simple modification of the standard guanidinium thiocyanate\u2010phenol\u2010chloroform extraction method, which involves enzymatic predigestion of the tissue prior to standard RNA isolation. Yield, purity and integrity of RNA isolated from bovine nucleus pulposus, inner annulus fibrosus and outer annulus fibrosus were compared among complete matrix digestion, predigestion and pulverization, pulverization alone, and pulverization followed by on\u2010column purification. With predigestion, the average yield of RNA obtained from bovine nucleus pulposus was 8.82\u2009\u00b1\u20092.05\u2009ng\/mg of wet tissue with 260\/280 and 260\/230 optical density ratios of 1.91\u2009\u00b1\u20090.15 and 1.84\u2009\u00b1\u20090.30, respectively. RIN analysis indicated that RNA quality was best preserved with the predigestion method (RNA integrity number\u2009&gt;\u20097), and the extracted RNA was suitable for real\u2010time polymerase chain reaction. This method is of importance for gene expression studies on intervertebral disc development, degeneration and repair, and we anticipate that it may be further applied to other tissues rich in proteoglycans.<\/jats:p>","DOI":"10.1002\/jsp2.1017","type":"journal-article","created":{"date-parts":[[2018,6,8]],"date-time":"2018-06-08T02:14:37Z","timestamp":1528424077000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":29,"title":["Isolation of high\u2010quality RNA from intervertebral disc tissue via pronase predigestion and tissue pulverization"],"prefix":"10.1002","volume":"1","author":[{"given":"Stephanie","family":"Caprez","sequence":"first","affiliation":[{"name":"AO Research Institute Davos  Davos Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ursula","family":"Menzel","sequence":"additional","affiliation":[{"name":"AO Research Institute Davos  Davos Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9754-6389","authenticated-orcid":false,"given":"Zhen","family":"Li","sequence":"additional","affiliation":[{"name":"AO Research Institute Davos  Davos Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9552-3653","authenticated-orcid":false,"given":"Sibylle","family":"Grad","sequence":"additional","affiliation":[{"name":"AO Research Institute Davos  Davos Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0262-1412","authenticated-orcid":false,"given":"Mauro","family":"Alini","sequence":"additional","affiliation":[{"name":"AO Research Institute Davos  Davos Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5458-3446","authenticated-orcid":false,"given":"Marianna","family":"Peroglio","sequence":"additional","affiliation":[{"name":"AO Research Institute Davos  Davos Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,6,19]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1097\/01.brs.0000231761.73859.2c"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.2200\/S00250ED1V01Y201006TIS005"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1089\/scd.2014.0203"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.spinee.2013.07.491"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2014.06.009"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.1988.68.3.858"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.2144\/97226bm16"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(92)90324-Z"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1006\/abio.2000.4779"},{"key":"e_1_2_6_11_1","first-page":"101","article-title":"RNA extraction from cartilage","volume":"100","author":"Mallein\u2010Gerin F","year":"2004","journal-title":"Methods Mol Med"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00586-007-0475-y"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2008.06.005"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1089\/ten.tec.2012.0693"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.14440\/jbm.2015.40"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1302\/2046-3758.511.BJR-2016-0033.R3"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(87)90021-2"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2199-7-3"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00586-011-1976-2"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00586-014-3198-x"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1097\/BRS.0b013e3181a55705"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1002\/jor.22942"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1093\/dnares\/dsn030"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1097\/BRS.0b013e318166e001"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1097\/BRS.0b013e31824d911c"}],"container-title":["JOR SPINE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjsp2.1017","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jsp2.1017","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jsp2.1017","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jsp2.1017","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,10]],"date-time":"2023-09-10T21:11:30Z","timestamp":1694380290000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jsp2.1017"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,6]]},"references-count":24,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2018,6]]}},"alternative-id":["10.1002\/jsp2.1017"],"URL":"https:\/\/doi.org\/10.1002\/jsp2.1017","archive":["Portico"],"relation":{},"ISSN":["2572-1143","2572-1143"],"issn-type":[{"value":"2572-1143","type":"print"},{"value":"2572-1143","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,6]]},"assertion":[{"value":"2018-04-09","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2018-05-27","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2018-06-19","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e1017"}}