Pig Jejunal Transcriptome Analysis Protocol

The pig jejunal explants were homogenized using 2 mL plastic bead tubes (MT Biomedicals, Illkirch, France) in 1 mL of Extract-All reagent (Eurobio, Courtaboeuf, France) in a Precellys Evolution tissue homogenizer (Bertin Technologies, Montigny-le-Bretonneux, France). ARN isolation and RT-qPCR were performed as described elsewhere [70 (link),71 (link)]. Data analysis was carried out using LinRegPCR freeware [72 (link)], and normalized against two reference genes, TATA-Box Binding Protein (TBP) and Hypoxanthine guanine phosphoribosyl transferase 1 (HPRT1). Primers are described in Supplementary Table S2.

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Lahjouji T., Bertaccini A., Neves M., Puel S., Oswald I.P, & Soler L. (2020). Acute Exposure to Zearalenone Disturbs Intestinal Homeostasis by Modulating the Wnt/β-Catenin Signaling Pathway. Toxins, 12(2), 113.

Publication 2020

Extract-All reagent Precellys Evolution tissue homogenizer

Evolution Genes Hypoxanthine guanine phosphoribosyl transferase Isolation Jejunal Primers Tata box binding protein Tissue

Corresponding Organization :

Other organizations : Toxalim Research Centre in Food Toxicology

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Variable analysis

independent variables

Homogenization method: 2 mL plastic bead tubes, Precellys Evolution tissue homogenizer

dependent variables

Gene expression levels (normalized against reference genes TBP and HPRT1)

control variables

Tissue: Pig jejunal explants
Reagent for homogenization: Extract-All reagent
Methods for RNA isolation and RT-qPCR: As described elsewhere [70, 71]
Data analysis: LinRegPCR freeware [72]
Reference genes for normalization: TBP and HPRT1

controls

No positive or negative controls were explicitly mentioned in the provided information.

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