Quantitative PCR Analysis of Gene Expression

Total RNA from participants with three (that is, BL, D2 and D6) or two (that is, either combination of BL, D2 or D6) matched samples were selected for quantitative real-time PCR analysis. Missing samples were due to either participant failing to attend one of the three clinic visits or sample RNA quantity being minimal. Genes were selected for expression confirmation based on PC1 loadings. Depending on the stage of ARI analyzed, the highest ranked (peak, D2 of reduction and resolution) or lowest (D6 of reduction) genes were selected. Gene expression in each sample was normalized to glyceraldehyde-3-phosphate dehydrogenase and peptidylprolyl isomerase A as outlined previously [32 (link)]. Significance of fold change in gene copy number between groups was determined by Mann Whitney U-test. Primer and probe sequences are available upon request.

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McErlean P., Berdnikovs S., Favoreto S J.r., Shen J., Biyasheva A., Barbeau R., Eisley C., Barczak A., Ward T., Schleimer R.P., Erle D.J., Boushey H.A, & Avila P.C. (2014). Asthmatics with exacerbation during acute respiratory illness exhibit unique transcriptional signatures within the nasal mucosa. Genome Medicine, 6(1), 1.

Publication 2014

Clinic visits Gene expression Genes Glyceraldehyde 3 phosphate dehydrogenase Peptidylprolyl isomerase Primer Quantitative real time pcr analysis

Corresponding Organization :

Other organizations : Northwestern University, University of California, San Francisco

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Variable analysis

independent variables

Time point of sample collection (BL, D2, D6)

dependent variables

Gene expression levels of selected genes

control variables

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and peptidylprolyl isomerase A (PPIA) as reference genes for normalization of gene expression

controls

Positive control: None specified
Negative control: None specified

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