Relative Gene Expression Analysis

The results are shown as the arithmetic means ±SD from three independent biological replicates. The results were evaluated with two-way analysis of variance (ANOVA) (factors: infection and duration of infection). A post hoc Tukey’s test was implemented to determine significant differences between means at p < 0.05 (Statistica 13.3; StatSoft Inc., Tulsa, OK, USA). The gene transcript level of each gene was tested in three biological replicates and three technical repetitions. The specificity of amplified PCR products was verified by melting curve analysis. For statistical analysis, the calculation of reaction efficiency was performed using LinRegPCR software version 2020.0.0.0.3 (http://LinRegPCR.nl), whereas the relative gene expression levels and statistical significance of their differences were estimated by Relative Expression Software Tool 2009 (REST 2009) (http://rest.gene-quantification.info/) as described previously [30 (link)] and confirmed using Student’s t-test and Fisher’s F-test (Statistica 13.3; StatSoft Inc.).

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Labudda M., Różańska E., Gietler M., Fidler J., Muszyńska E., Prabucka B, & Morkunas I. (2020). Cyst Nematode Infection Elicits Alteration in the Level of Reactive Nitrogen Species, Protein S-Nitrosylation and Nitration, and Nitrosoglutathione Reductase in Arabidopsis thaliana Roots. Antioxidants, 9(9), 795.

Publication 2020

Statistica 13.3

Biological Gene Gene expression Infection Student

Corresponding Organization : Warsaw University of Life Sciences

Other organizations : University of Life Sciences in Poznań

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Variable analysis

independent variables

Infection
Duration of infection

dependent variables

Gene transcript level of each gene

control variables

Three biological replicates and three technical repetitions for gene transcript level testing
Specificity of amplified PCR products verified by melting curve analysis

controls

Positive controls not explicitly mentioned
Negative controls not explicitly mentioned

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