Hello,
I'm relatively new to qPCR and have been utilizing a manufacturer's analysis software for my experiments, including one-way ANOVA for statistical analysis. However, I've encountered a discrepancy when manually conducting the same statistical analysis using CT values imported from the software (because i'd like to conduct some post-hoc tests too). Specifically, I've noticed that genes with higher CT values appear to be more expressed when considering dCt and ddCt values calculated within the app. Could you tell me why this difference occurs, especially given my beginner level of experience with qPCR?
Dear Dominika Celar
ensure consistency in data processing steps, normalization methods, and statistical analysis techniques between the manufacturer's software and your manual analysis. Double-check the settings and parameters used in both analyses to identify any potential differences that may be causing the discrepancy. Additionally, consulting with experienced colleagues or seeking guidance from experts in qPCR analysis can help troubleshoot the issue and ensure the accuracy of your results.
It sounds like you're experiencing a discrepancy between the results obtained from the manufacturer's analysis software and those obtained when conducting your statistical analysis manually using CT values. This can be a common issue, especially when you're new to qPCR analysis.
When conducting statistical analysis, such as one-way ANOVA and post-hoc tests, ensure that you're using the appropriate statistical methods and assumptions. Different methods may yield slightly different results, especially if assumptions are violated or if there are discrepancies in the data input.
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