Frequently Asked Questions

Yes, completely free. There are no hidden fees, no premium tiers, no feature locks, and no time limits. All features are available to everyone. The site is supported by advertising.

Your data is never stored on our servers. Files you upload are processed to generate your results, but no data is retained after your session. qPCR Guru does not save, share, or sell your experimental data. This makes it suitable for sensitive or proprietary research data.

qPCR Guru accepts XLS, XLSX, CSV, TXT, TSV, and RDML files. It auto-detects exports from QuantStudio, StepOne, ViiA 7, Bio-Rad CFX, Roche LightCycler, Agilent AriaMx, Qiagen Rotor-Gene, Eppendorf realplex, and generic tabular formats. You can also upload multiple files simultaneously for multi-plate analysis.

MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) is a set of guidelines published by Bustin et al. to ensure the quality and reproducibility of qPCR data. MIQE 2.0 (2025) is the latest update. qPCR Guru checks your data against MIQE 2.0 essential criteria including NTC validation, replicate consistency, reference gene stability, and efficiency correction — and shows you a real-time compliance score.

The ΔΔCq (also written ΔΔCt) method calculates relative gene expression by normalizing each target gene to a reference gene (ΔCq) and then comparing to a calibrator sample (ΔΔCq). The relative quantity (RQ) is calculated as 2^-ΔΔCq. When primer efficiency data is available, qPCR Guru uses the more accurate Pfaffl method, which accounts for differing PCR efficiencies between target and reference genes.

qPCR Guru offers Welch's t-test (two groups, parametric), Mann-Whitney U (two groups, non-parametric), one-way ANOVA with Tukey HSD post-hoc (3+ groups, parametric), and Kruskal-Wallis (3+ groups, non-parametric). An "Auto-select" mode intelligently chooses the appropriate test based on your number of groups. Benjamini-Hochberg FDR correction is available for multiple testing across targets.

Absolutely. All figures can be exported at 300+ DPI in JPEG, TIFF, or PDF format — meeting the resolution requirements of all major journals. The MIQE 2.0 compliance panel helps ensure your analysis meets the reporting standards that reviewers expect. You can also generate a comprehensive multi-page PDF report suitable for supplementary materials.

geNorm M (Vandesompele et al., 2002) evaluates reference gene stability by computing pairwise variation across all candidate reference genes. NormFinder ρ (Andersen et al., 2004) estimates stability by measuring the variation of each candidate gene across samples independently. When both algorithms agree on the most stable gene, you have high confidence. When they disagree, it flags potential issues worth investigating.

RDML (Real-Time PCR Data Markup Language) is the standardized XML format for qPCR data exchange, endorsed by the MIQE guidelines. It allows you to share your raw data with collaborators regardless of what instrument or software they use. qPCR Guru can both import RDML files (including ZIP-compressed ones) and export your data as RDML for archival or sharing.

Use the Sample Groups feature in the ΔΔCq Analysis tab. Group your independent biological samples (e.g., different mice, patients) under a single condition name. Technical replicates (same cDNA in multiple wells) are automatically averaged within each sample before group-level statistics are calculated. This ensures your p-values reflect biological variation, not pipetting precision — a key MIQE 2.0 requirement.

We provide a comprehensive User Guide (PDF) covering every feature of qPCR Guru — from uploading your data and running QC, through setting up your analysis, interpreting charts, running statistical tests, and exporting publication-ready figures. The guide is also accessible from the upload screen and the navigation bar on every page.