Outside the lines

Unlock the Power of qPCR

Quantitative Polymerase Chain Reaction (qPCR) is a precise molecular biology technique enabling the accurate quantification of DNA sequences, crucial for gene expression and target presence analysis in biological samples. With real-time monitoring capabilities and a broad dynamic range, qPCR is an invaluable tool for numerous research applications.

Target Audience

This training, open to seasoned researchers, PhD students, and industry professionals, builds on Bram's +10 years of experience in training people in Belgium and abroad. Enhance your qPCR skills and stay at the forefront of molecular biology by joining this course!

The complete workflow explained:

1. Experiment design

Power analysis
Experiment layout (sample vs gene maximization)
Biological and technical replicates
Inter-run calibration

2. Sample preparation and quality control

RNA integrity and purity
DNase treatment
cDNA synthesis
Pre-amplification

3. Assay design and validation

Primer design
Probes vs intercalating dye
Design guidelines
In silico evaluation
Empirical validation

4. qPCR amplification

Real-time PCR principle
Amplification curve
Melt curve
Cq value determination methods
Replicates and controls
Speed and throughput considerations

5. Absolute quantification

Standard curves
Limitations and concerns

6. Relative quantification

Quantification models
Efficiency correction
Selection and validation of reference genes
Normalization with multiple reference genes
Alternative normalization methods
Inter-run calibration
Result rescaling

7. Data quality and control

Visual inspection of plots
Positive and negative controls
Normalization factor histogram
Reference gene stability
Inter-run variation

8. Bio-statistical analysis

Basic principles
Descriptive statistics
Selection and application of appropriate statistical tests

Included

Electronic version of the course is provided 1 week before the start of the course.

Certificate of attendance will be provided after the course