Filll - iPCR - Tool for Interactive qPCR Data Analysis

Preprocessing and Analysis
ΔΔCq Analysis
ΔCq ANOVA (Dunnett’s post-hoc)
ΔΔCq ANOVA (Dunnett’s post-hoc)
Diagnostics

Template

Upload Your Data CSV File:

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Enable multiple reference genes

3. Comparison Settings

Color Palette:

Download Stats

Download Plot

Save Displayed Size

Download Plot Settings

Settings for the 'Download Plot' button.

Width (inches):

Height (inches):

Fix the ratio

Resolution (DPI):

1. ΔCq Calculation:
ΔCq = Cq(Target Gene) - Mean Cq(Reference Genes)

2. Statistical Test:
A Welch's two-sample t-test is performed on the ΔCq values between each treatment group and the control group to calculate the p-value.

3. Visualization:
The plot displays Relative Expression (2^-ΔCq) for intuitive visualization.

Reference Gene(s):

ΔΔCq Comparison Settings

Color Palette:

Download ΔΔCq Stats

Download ΔΔCq Plot

Save Displayed Size

Download Plot Settings

Settings for the 'Download ΔΔCq Plot' button.

Width (inches):

Height (inches):

Fix the ratio

Resolution (DPI):

Appendix: Statistical Method

1. ΔΔCq Calculation:
ΔΔCq = ΔCq(Sample) - Mean ΔCq(Control Group)
(where ΔCq = Cq_target - Mean Cq_refs)

2. Statistical Test:
A Welch's two-sample t-test is performed on the ΔCq values between each treatment group and the control group to calculate the p-value.

3. Visualization:
The plot displays Relative Expression (2^-ΔΔCq) for intuitive visualization.

This tab is for comparing 3 or more groups. It performs an ANOVA followed by Dunnett's test to compare each treatment group against a single control group.

Reference Gene(s):

ANOVA (3+ Groups) Comparison Settings

Color Palette:

Download ANOVA Stats

Download ANOVA Plot

Save Displayed Size

Download Plot Settings

Settings for the 'Download ANOVA Plot' button.

Width (inches):

Height (inches):

Fix the ratio

Resolution (DPI):

Appendix: Statistical Method

1. ΔCq Calculation:
ΔCq = Cq(Target Gene) - Mean Cq(Reference Genes)

2. Statistical Test:
First, a one-way ANOVA is performed on the ΔCq values to determine if there are any significant differences among the means of all selected groups. Then, Dunnett's test is used as a post-hoc test to specifically compare each treatment group against the single control group.

3. Visualization:
The plot displays Relative Expression (2^-ΔCq) for intuitive visualization.

This tab visualizes the ANOVA results as Fold Change (2^-ΔΔCq). The statistical test is the same as the 'ANOVA Analysis' tab.

Reference Gene(s):

ANOVA (ΔΔCq) Comparison Settings

Color Palette:

Download Stats

Download Plot

Save Displayed Size

Download Plot Settings

Settings for the 'Download Plot' button.

Width (inches):

Height (inches):

Fix the ratio

Resolution (DPI):

Appendix: Statistical Method

1. ΔΔCq Calculation:
ΔΔCq = ΔCq(Sample) - Mean ΔCq(Control Group)
(where ΔCq = Cq_target - Mean Cq_refs)

2. Statistical Test:
The statistical test is identical to the 'ANOVA Analysis' tab (ANOVA followed by Dunnett's test on ΔCq values).

3. Visualization:
The plot displays Fold Change (2^-ΔΔCq) to visualize expression relative to the control group.

App Functionality Diagnostics

Use this panel to verify that the app's core functions are working correctly with a sample dataset.

Notifications:
# Calculate the mean of Cq values by yourself, if you have technical replicates.
# Please cite this paper if you use this app in your research. A. Kubota and A. Tajima, bioRxiv, 2025. (https://doi.org/10.1101/2025.05.29.656779).

View User Guide (English and Japanese) & Source Code on GitHub