Data Source and Methodology
Authoritative data sourced from GoldBio qPCR Analysis Tool. All calculations are based strictly on the formulas and data provided by this source.
The Formula Explained
\[\Delta\Delta C_t = \Delta C_t (\text{Test}) - \Delta C_t (\text{Control})\]
\[\Delta C_t = C_t (\text{Target}) - C_t (\text{Reference})\]
Glossary of Variables
- Control Sample Ct: Ct value of the control sample.
- Test Sample Ct: Ct value of the test sample.
- Reference Gene Ct: Ct value of the reference gene.
- Fold Change: The calculated expression change relative to the control.
How It Works: A Step-by-Step Example
For example, if the Test Sample Ct is 25, the Control Sample Ct is 20, and the Reference Gene Ct is 18, the fold change is calculated as follows:
First, calculate the ΔCt for both the test and control samples, then find the ΔΔCt by subtracting the control ΔCt from the test ΔCt. Finally, calculate the fold change using the formula \(2^{-\Delta\Delta C_t}\).
Frequently Asked Questions (FAQ)
What is qPCR?
qPCR, or quantitative PCR, is a technique used to amplify and simultaneously quantify a targeted DNA molecule.
How do I interpret the fold change?
A fold change greater than 1 indicates an up-regulation, whereas less than 1 indicates a down-regulation.
What are Ct values?
Ct values, or cycle threshold values, represent the cycle number at which the fluorescence of a PCR product exceeds the background fluorescence.
Why is the reference gene important?
The reference gene is used to normalize the expression levels of the target gene.
Can I use this calculator for all experiments?
This calculator is designed for qPCR data analysis using the Delta-Delta Ct method, which is widely applicable in gene expression studies.