Quantitative Real-Time PCR Analysis of Gene Expression

Total RNA was extracted using Trizol Reagent (Invitrogen) and reverse-transcribed into cDNA using PrimeScript RT Master Mix (TaKaRa). Real-time PCR was performed using the TB Green Premix Ex Taq II system (TaKaRa) with an Applied Biosystem 7500 instrument (Applied Biosystems).21 (link) Primer sequences were listed in online supplemental table 1. Relative expression to β-actin was determined using the ∆∆Ct method.

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Yang Y., Li L., Xu C., Wang Y., Wang Z., Chen M., Jiang Z., Pan J., Yang C., Li X., Song K., Yan J., Xie W., Wu X., Chen Z., Yuan Y., Zheng S., Yan J., Huang J, & Qiu F. (2020). Cross-talk between the gut microbiota and monocyte-like macrophages mediates an inflammatory response to promote colitis-associated tumourigenesis. Gut, 70(8), 1495-1506.

Publication 2020

Trizol Reagent PrimeScript RT Master Mix TB Green Premix Ex Taq II system

Actin Cdna Primer Real time pcr Trizol

Corresponding Organization : Second Affiliated Hospital of Zhejiang University

Other organizations : Zhejiang University, Zhejiang Hospital, Zhuhai People's Hospital, University of Louisville

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Relative expression to β-actin

control variables

RNA extraction using Trizol Reagent (Invitrogen)
Reverse transcription into cDNA using PrimeScript RT Master Mix (TaKaRa)
Real-time PCR using TB Green Premix Ex Taq II system (TaKaRa) with Applied Biosystem 7500 instrument (Applied Biosystems)
Primer sequences listed in online supplemental table 1

controls

Positive control: Not mentioned
Negative control: Not mentioned

Annotations

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