Quantitative Analysis of Respiratory Genes

Messenger RNA (mRNA) was isolated using TRIzol as previously described (39 (link)), and complementary DNA (cDNA) was generated from DNase-treated mRNA using a reverse transcription kit (Life Technologies). Quantitative PCR was performed using EvaGreen master mix (BioRad) or TaqMan primers and universal master mix (Life Technologies) on the Mx3005P system (Agilent Technologies). The primers for il13, gob5, and RSV Large polymerase (L) gene (31 (link)) were purchased from IDT and relative gene expression was determined as 2^−ΔΔCT (39 (link)).

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Krishnamoorthy N., Walker K.H., Brüggemann T.R., Tavares L.P., Smith E.W., Nijmeh J., Bai Y., Ai X., Cagnina R.E., Duvall M.G., Lehoczky J.A, & Levy B.D. (2023). The Maresin 1–LGR6 axis decreases respiratory syncytial virus-induced lung inflammation. Proceedings of the National Academy of Sciences of the United States of America, 120(2), e2206480120.

Publication 2023

reverse transcription kit EvaGreen master mix TaqMan primers universal master mix Mx3005P system

Cdna Dnase Gene Gene expression Il13 Messenger rna Primers Reverse transcription Trizol

Corresponding Organization :

Other organizations : Harvard University, Brigham and Women's Hospital, Massachusetts General Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression of il13, gob5, and RSV Large polymerase (L) gene

control variables

None explicitly mentioned

controls

Positive control: None specified
Negative control: None specified

Annotations

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