Quantitative Real-Time PCR Analysis of Intestinal Gene Expression

Quantitative real-time PCR (RT-qPCR) analysis was performed, as described previously [20 (link)]. Briefly, the total RNA of the duodenum, jejunum, and ileum was extracted using the TRIZOL reagent (Invitrogen, Carlsbad, CA, USA) and then reversely transcribed into complementary DNA (cDNA) using RevertAid reverse transcriptase (Takara, Otsu, Japan). RT-qPCR analysis was performed with SYBR green I fluorescent dye (Takara, Otsu, Japan). The relative levels of the mRNA expression of the target genes were calculated using the 2^−ΔΔCt method [21 (link),22 (link)]. All primer sequences are presented in Table 2.

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Liu F., Li J., Ni H., Azad M.A., Mo K, & Yin Y. (2023). The Effects of Phytase and Non-Starch Polysaccharide-Hydrolyzing Enzymes on Trace Element Deposition, Intestinal Morphology, and Cecal Microbiota of Growing–Finishing Pigs. Animals : an Open Access Journal from MDPI, 13(4), 549.

Publication 2023

TRIZOL reagent SYBR green I fluorescent dye

Cdna Duodenum Expression genes Fluorescent dye Ileum Jejunum Mrna Primer Quantitative real time pcr analysis Real time pcr analysis Reverse transcriptase Sybr green i Trizol

Corresponding Organization : Institute of Subtropical Agriculture

Other organizations : Hunan Agricultural University, South China Agricultural University

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Variable analysis

independent variables

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dependent variables

Relative levels of mRNA expression of target genes

control variables

None explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

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