Quantification of Gene Expression by qPCR

It was performed as described previously [24 (link)]. Briefly, total mRNA from cells was collected using TRI reagent (#TR118; Molecular Research Center, Cincinnati, OH, USA). cDNA was synthesized using the Superscript Reverse Transcription system (Takara, Shiga, Japan), and real-time quantitative PCR (qPCR) was performed using LightCycler FastStart DNA Master SYBR Green I (Takara) in accordance with the manufacturer’s instructions. Expression levels were normalized relative to β-actin. The sequences of the primers used are listed in Additional file 1: Table S1.

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Lee A.R., Woo J.S., Lee S.Y., Lee Y.S., Jung J., Lee C.R., Park S.H, & Cho M.L. (2023). SARS-CoV-2 spike protein promotes inflammatory cytokine activation and aggravates rheumatoid arthritis. Cell Communication and Signaling : CCS, 21, 44.

Publication 2023

TRI reagent Superscript Reverse Transcription system LightCycler FastStart DNA Master SYBR Green I

Actin Cdna Cells Mrna Primers Real time pcr Reverse transcription Sybr green i

Corresponding Organization : Seoul St. Mary's Hospital

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Variable analysis

independent variables

MRNA collection using TRI reagent
CDNA synthesis using Superscript Reverse Transcription system
Real-time quantitative PCR (qPCR) using LightCycler FastStart DNA Master SYBR Green I

dependent variables

Expression levels of target genes

control variables

β-actin as the reference gene for normalization

controls

No positive or negative controls were explicitly mentioned.

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