Experimental Design and Statistical Data Analysis for qPCR

06 Sep 2012 - 07 Sep 2012, at 09:00-17:00 in Frankfurt, Germany

2 days Experimental Design and Statistical Data Analysis for qPCR

Course Description

Measuring qPCR is easy; reaching biologically relevant conclusions from RT-qPCR studies is much more difficult. Natural variation between subjects and technical noise introduced by the experimental procedure confounds the effect of treatment. Qualified qPCR measurements require controlling the confounding variation using appropriate controls and references and separating the noise from the biological variation or treatment effect. In this course you will learn the basics of statistics as applied to qPCR, how to design qPCR experiments, appropriate handling and pre-processing of qPCR data for analysis, absolute quantification, relative quantification and expression profiling.  

Target Audience

This course is intended for researchers and laboratory personnel working with or planning to work with real-time PCR, researchers with interest in gene and protein expression, clinicians and pathologists with interest in molecular diagnostics, virologists, bacteriologists, and scientists with interest in food diagnostics and environmental testing. 

Level of the Course

From beginners to advanced users

Course Outline 

Day 1 - Statistical analysis of real-time PCR data 

• Principles of statistics

• Descriptive statistics

• Normal distribution, null hypothesis, p-values

• T-test, non-parametric tests, ANOVA

• Power of a test (ability to detect a difference)

• qPCR data pre-processing

• Relative quantification (group comparison)

Day 2 - Gene expression profiling with real-time PCR

• Multiplate measurements, Interplate calibration

• Standard curves, absolute quantification (comparison with standards)

• Limit of detection, limit of quantification

• Experimental design, performance and cost optimization

• Normalization, reference gene selection and validation

• Expression profiling

• Hierarchical clustering, heat map

• Principal component analysis, Self organized maps

Instructor Biography

Professor Mikael Kubista was among the pioneers who developed real-time PCR. In 1991 his laboratory developed dyes and probes for qPCR and founded LightUp Technologies as the first company focusing on qPCR based human infectious disease diagnostics. His team developed methods and approaches to analyze qPCR data and founded MultiD Analyses (www.multid.se) which develops market leading GenEx software for qPCR experimental design and data analysis. Kubista’s team also pioneered single cell expression profiling, multidimensional expression profiling and intracellular expression profiling by qPCR. As advisor to Unesco Kubista introduced qPCR in Africa and in the Middle East. In 2001 Kubista founded the TATAA Biocenters (www.tataa.com), which are Europe’s leading qPCR service provider and organizer of hands-on training workshops. The TATAA courses are supported world-wide by leading instrument manufacturers and reagent suppliers in the field. TATAA Biocenter is member of SPIDIA (www.spidia.eu), which consortium is developing guidelines for pre-analytics in molecular diagnostics. The TATAA Biocenters also organize the main annual QPCR symposium (www.qpcrsymposium.eu). Kubista is advisor to several leading companies in the qPCR field including Life Technologies and Roche. He is editorial board member of Scientific Reports (Nature Publishing Group).