{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T13:49:31Z","timestamp":1779889771063,"version":"3.53.1"},"reference-count":37,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T00:00:00Z","timestamp":1748908800000},"content-version":"vor","delay-in-days":153,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,1,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Gene expression analysis via reverse transcription quantitative real-time polymerase chain reaction (qPCR) can be inhibited by various substances, including dextran sodium sulfate (DSS), a chemical commonly used to induce intestinal inflammation in animal models. Ensuring elimination and reduction of qPCR interference in tissues from laboratory animals following oral administration of DSS is critical and may improve the power of experimental tests. While methods for DSS removal have been reported, their effectiveness varies depending on the animal model, extraction techniques, DSS concentration, and treatment duration. We compared the effectiveness of the commonly used RNeasy Plus Universal Mini Kit with and without further lithium chloride (LiCl) precipitation at eliminating or reducing qPCR interference from RNA isolated from the colons of DSS-treated mice with histologically confirmed intestinal damage. The RNeasy Plus Universal Mini Kit alone was insufficient to eliminate interference in colonic tissue, as evidenced by increased quantification cycle (Cq) values and variation in the reference gene expression in DSS-treated distal colons (P adj.\u2005=\u20050.04). LiCl precipitation restored Cq values to control levels and reduced variation. DSS treatment did not lead to interference in non-enteric tissues, including the spleen and cortex. LiCl precipitation not only restored reference gene expression but also improved the detection of changes in inflammatory markers, Il-6 and Tnf, in colonic tissue. In proximal colons, Il-6 was upregulated 2.97-fold in DSS-treated (non-LiCl precipitated, P adj.\u2005=\u20050.007), and 4.00-fold following LiCl precipitation (P adj.\u2005=\u20050.0004), compared to control mice. In distal colons, Il-6 was upregulated 3.95-fold in DSS-treated (non-LiCl precipitated, P adj.\u2005=\u20050.014) and 5.57-fold after LiCl precipitation (P adj.\u2005=\u20050.001) compared to control. Tnf was upregulated 2.05-fold in DSS-treated (non-LiCl precipitated, P adj.\u2005=\u20050.04), and 2.44 (P adj.\u2005=\u20050.009) after LiCl precipitation compared to controls in proximal colons. In distal colons, Tnf was 3.47-fold higher in DSS-treated (non-LiCl precipitated, P adj.\u2005=\u20050.009) and 4.41-fold higher after LiCl precipitation (P adj.\u2005=\u20050.002) compared to control. These findings demonstrate that the combined use of the RNeasy Plus Universal Mini Kit and LiCl precipitation enhances qPCR performance, ensuring reliable gene expression analysis in colonic tissue following acute DSS treatment in a murine model.<\/jats:p>","DOI":"10.1093\/jas\/skaf188","type":"journal-article","created":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T04:32:32Z","timestamp":1748925152000},"source":"Crossref","is-referenced-by-count":2,"title":["Reducing dextran sodium sulfate interference with gene expression quantification in a mouse model of colitis"],"prefix":"10.1093","volume":"103","author":[{"given":"Drake","family":"Hechter","sequence":"first","affiliation":[{"name":"Department of Food and Human Nutritional Sciences, University of Manitoba , Winnipeg, MB , R3T 2N2","place":["Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0563-7724","authenticated-orcid":false,"given":"Sara V","family":"Good","sequence":"additional","affiliation":[{"name":"Department of Food and Human Nutritional Sciences, University of Manitoba , Winnipeg, MB , R3T 2N2","place":["Canada"]},{"name":"Department of Biology, The University of Winnipeg , Winnipeg, MB , R3B 2E9","place":["Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,6,3]]},"reference":[{"key":"2025072407570472100_CIT0001","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1007\/978-1-59745-447-6_5","article-title":"Approaches to determine expression of inflammatory cytokines","volume-title":"Inflammation and cancer, methods in molecular biology","author":"Amsen","year":"2009"},{"key":"2025072407570472100_CIT0002","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1007\/s10620-011-1881-x","article-title":"The severity of dextran sodium sulfate-induced colitis can differ between dextran sodium sulfate preparations of the same molecular weight range","volume":"57","author":"Bamba","year":"2012","journal-title":"Dig. Dis. Sci"},{"key":"2025072407570472100_CIT0003","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1096\/fba.2019-00035","article-title":"Weight loss is a sufficient and economical single outcome measure of murine dextran sulfate sodium colitis","volume":"1","author":"Britto","year":"2019","journal-title":"FASEB Bioadv"},{"key":"2025072407570472100_CIT0005","first-page":"155","article-title":"Pitfalls of quantitative real-time reverse-transcription polymerase chain reaction","volume":"15","author":"Bustin","year":"2004","journal-title":"J. Biomol. Tech"},{"key":"2025072407570472100_CIT0004","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin. Chem"},{"key":"2025072407570472100_CIT0006","doi-asserted-by":"publisher","DOI":"10.1002\/0471142735.im1525s104","article-title":"Dextran Sulfate Sodium (DSS)-induced colitis in mice","volume":"104:15.25","author":"Chassaing","year":"2014","journal-title":"Curr. Protoc. Immunol"},{"key":"2025072407570472100_CIT0008","first-page":"4557","article-title":"A guide to histomorphological evaluation of intestinal inflammation in mouse models","author":"Erben","year":"2014","journal-title":"Int. J. Clin. Exp. Path"},{"key":"2025072407570472100_CIT0007","doi-asserted-by":"publisher","first-page":"1","DOI":"10.14440\/jbm.2017.184","article-title":"A cost-effective RNA extraction technique from animal cells and tissue using silica columns","volume":"4;7","author":"Escobar","year":"2017","journal-title":"JBM"},{"key":"2025072407570472100_CIT0009","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1016\/j.mam.2005.12.003","article-title":"RNA integrity and the effect on the real-time qRT-PCR performance","volume":"27","author":"Fleige","year":"2006","journal-title":"Mol. Aspects Med"},{"key":"2025072407570472100_CIT0010","doi-asserted-by":"publisher","first-page":"650","DOI":"10.1111\/j.1755-0998.2011.03010.x","article-title":"Next-generation sequencing of transcriptomes: a guide to RNA isolation in nonmodel animals","volume":"11","author":"Gayral","year":"2011","journal-title":"Mol. Ecol. Resour"},{"key":"2025072407570472100_CIT0011","doi-asserted-by":"publisher","first-page":"e0182732","DOI":"10.1371\/journal.pone.0182732","article-title":"Dextran sodium sulfate (DSS) induces necrotizing enterocolitis-like lesions in neonatal mice","volume":"12","author":"Ginzel","year":"2017","journal-title":"PLoS One"},{"key":"2025072407570472100_CIT0012","doi-asserted-by":"publisher","first-page":"386","DOI":"10.1006\/meth.2001.1261","article-title":"An overview of real-time quantitative PCR: applications to quantify cytokine gene expression","volume":"25","author":"Giulietti","year":"2001","journal-title":"Methods"},{"key":"2025072407570472100_CIT0013","article-title":"Caloric restriction alters colonic and splenic immune responses and microbial composition in a murine model of acute DSS-Induced colitis. Submitted","author":"Hechter","year":"2025"},{"key":"2025072407570472100_CIT0014","doi-asserted-by":"publisher","first-page":"e69983","DOI":"10.1371\/journal.pone.0069983","article-title":"Soybean meal induces intestinal inflammation in zebrafish larvae","volume":"8","author":"Hedrera","year":"2013","journal-title":"PLoS One"},{"key":"2025072407570472100_CIT0015","doi-asserted-by":"publisher","first-page":"255","DOI":"10.1128\/JVI.27.1.255-257.1978","article-title":"Dextran sulfates as a contaminant of DNA extracted from concentrated viruses and as an inhibitor of DNA polymerases","volume":"27","author":"Hitzeman","year":"1978","journal-title":"J. Virol"},{"key":"2025072407570472100_CIT0016","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1016\/j.ab.2019.05.011","article-title":"Rapid removal of dextran sulfate sodium from tissue RNA preparations for measurement of inflammation biomarkers","volume":"579","author":"Juritsch","year":"2019","journal-title":"Anal. Biochem"},{"key":"2025072407570472100_CIT0017","doi-asserted-by":"publisher","first-page":"344","DOI":"10.1002\/ibd.21763","article-title":"Dextran sodium sulfate inhibition of real-time polymerase chain reaction amplification: a poly-A purification solution","volume":"18","author":"Kerr","year":"2012","journal-title":"Inflamm. Bowel Dis"},{"key":"2025072407570472100_CIT0018","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.mimet.2017.10.015","article-title":"Have you tried spermine? A rapid and cost-effective method to eliminate dextran sodium sulfate inhibition of PCR and RT-PCR","volume":"144","author":"Krych","year":"2018","journal-title":"J. Microbiol. Methods"},{"key":"2025072407570472100_CIT0019","doi-asserted-by":"publisher","first-page":"5283","DOI":"10.1038\/s41598-021-84761-1","article-title":"DSS-induced colitis is associated with adipose tissue dysfunction and disrupted hepatic lipid metabolism leading to hepatosteatosis and dyslipidemia in mice","volume":"11","author":"Kwon","year":"2021","journal-title":"Sci. Rep"},{"key":"2025072407570472100_CIT0020","doi-asserted-by":"publisher","first-page":"447","DOI":"10.1242\/dmm.007252","article-title":"Worms, flies and four-legged friends: the applicability of biological models to the understanding of intestinal inflammatory diseases","volume":"4","author":"Lin","year":"2011","journal-title":"Dis Model Mech"},{"key":"2025072407570472100_CIT0021","doi-asserted-by":"publisher","first-page":"e0190846","DOI":"10.1371\/journal.pone.0190846","article-title":"Impaired resolution of DSS-induced colitis in mice lacking the glucocorticoid receptor in myeloid cells","volume":"13","author":"Meers","year":"2018","journal-title":"PLoS One"},{"key":"2025072407570472100_CIT0022","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1038\/nri3661","article-title":"Cytokines in inflammatory bowel disease","volume":"14","author":"Neurath","year":"2014","journal-title":"Nat. Rev. Immunol"},{"key":"2025072407570472100_CIT0023","doi-asserted-by":"publisher","first-page":"1327","DOI":"10.11405\/nisshoshi1964.82.1327","article-title":"Production of experimental ulcerative colitis in hamsters by dextran sulfate sodium and change in intestinal microflora","author":"Ohkusa","year":"1985"},{"key":"2025072407570472100_CIT0024","doi-asserted-by":"publisher","first-page":"694","DOI":"10.1016\/0016-5085(90)90290-h","article-title":"A novel method in the induction of reliable experimental acute and chronic ulcerative colitis in mice","volume":"98","author":"Okayasu","year":"1990","journal-title":"Gastroenterol"},{"key":"2025072407570472100_CIT0025","doi-asserted-by":"publisher","first-page":"61","DOI":"10.7171\/jbt.18-2903-001","article-title":"RNA purity, real-time PCR sensitivity, and colon segment influence mRNA relative expression in murine dextran sodium sulfate experimental colitis","volume":"29","author":"Oldak","year":"2018","journal-title":"J. Biomol. Tech"},{"key":"2025072407570472100_CIT0026","doi-asserted-by":"publisher","first-page":"2704","DOI":"10.1016\/j.celrep.2019.02.019","article-title":"Fasting-Mimicking diet modulates microbiota and promotes intestinal regeneration to reduce inflammatory bowel disease pathology","volume":"26","author":"Rangan","year":"2019","journal-title":"Cell Rep"},{"key":"2025072407570472100_CIT0027","doi-asserted-by":"publisher","first-page":"1101","DOI":"10.1038\/nprot.2008.73","article-title":"Analyzing real-time PCR data by the comparative CT method","volume":"3","author":"Schmittgen","year":"2008","journal-title":"Nat. Protoc"},{"key":"2025072407570472100_CIT0028","doi-asserted-by":"publisher","first-page":"skad202","DOI":"10.1093\/jas\/skad202","article-title":"Purification methods to reduce interference by dextran sodium sulfate with quantification of gene expression in intestinal tissue samples from a piglet model of colitis","volume":"101","author":"Sommer","year":"2023","journal-title":"J. Anim. Sci"},{"key":"2025072407570472100_CIT0029","article-title":"An experimental model of colitis induced by dextran sulfate sodium from acute progresses to chronicity in C57BL\/6: correlation between conditions of mice and","author":"Taghipour","year":"2016"},{"key":"2025072407570472100_CIT0030","doi-asserted-by":"publisher","first-page":"3572","DOI":"10.3748\/wjg.v25.i27.3572","article-title":"Mucosal healing progression after acute colitis in mice","volume":"25","author":"Vidal-Lletj\u00f3s","year":"2019","journal-title":"World J. Gastroenterol"},{"key":"2025072407570472100_CIT0031","doi-asserted-by":"publisher","first-page":"360","DOI":"10.1186\/1756-0500-6-360","article-title":"Dextran sodium sulfate inhibits the activities of both polymerase and reverse transcriptase: lithium chloride purification, a rapid and efficient technique to purify RNA","volume":"6","author":"Viennois","year":"2013","journal-title":"BMC Res Notes"},{"key":"2025072407570472100_CIT0032","first-page":"266","article-title":"Analysis of one-step and two-step real-time RT-PCR using superscript III","volume":"16","author":"Wacker","year":"2005","journal-title":"J. Biomol. Tech"},{"key":"2025072407570472100_CIT0033","doi-asserted-by":"publisher","first-page":"541","DOI":"10.1038\/nprot.2007.41","article-title":"Chemically induced mouse models of intestinal inflammation","volume":"2","author":"Wirtz","year":"2007","journal-title":"Nat. Protoc"},{"key":"2025072407570472100_CIT0034","doi-asserted-by":"publisher","first-page":"5311","DOI":"10.3390\/nu14245311","article-title":"Intermittent fasting alleviates risk markers in a murine model of ulcerative colitis by modulating the gut microbiome and metabolome","volume":"14","author":"Wu","year":"2022","journal-title":"Nutrients"},{"key":"2025072407570472100_CIT0035","doi-asserted-by":"publisher","first-page":"844","DOI":"10.1093\/ibd\/izad312","article-title":"Unveiling colitis: A journey through the dextran sodium sulfate-induced model","volume":"30","author":"Yang","year":"2024","journal-title":"Inflamm. Bowel Dis"},{"key":"2025072407570472100_CIT0036","doi-asserted-by":"publisher","first-page":"101535","DOI":"10.1016\/j.redox.2020.101535","article-title":"Effects of alternate-day fasting, time-restricted fasting and intermittent energy restriction DSS-induced on colitis and behavioral disorders","volume":"32","author":"Zhang","year":"2020","journal-title":"Redox Biol"},{"key":"2025072407570472100_CIT0037","doi-asserted-by":"publisher","first-page":"25","DOI":"10.15171\/bi.2016.04","article-title":"A microRNA isolation method from clinical samples","volume":"6","author":"Zununi Vahed","year":"2016","journal-title":"Bioimpacts"}],"container-title":["Journal of Animal Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jas\/advance-article-pdf\/doi\/10.1093\/jas\/skaf188\/63422252\/skaf188.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jas\/article-pdf\/doi\/10.1093\/jas\/skaf188\/63422252\/skaf188.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jas\/article-pdf\/doi\/10.1093\/jas\/skaf188\/63422252\/skaf188.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,24]],"date-time":"2025-07-24T11:57:12Z","timestamp":1753358232000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jas\/article\/doi\/10.1093\/jas\/skaf188\/8155668"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025]]},"references-count":37,"URL":"https:\/\/doi.org\/10.1093\/jas\/skaf188","relation":{},"ISSN":["0021-8812","1525-3163"],"issn-type":[{"value":"0021-8812","type":"print"},{"value":"1525-3163","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2025]]},"published":{"date-parts":[[2025]]},"article-number":"skaf188"}}