Course Information
Course Title: Quality Assurance in Real-time PCR
Categories: 1 - Life Sciences
2 - Pharmaceutical Sciences
3 - Clinical Analysis
4 - Polymerase Chain Reaction
5 - Quality/Regulatory/Compliance
6 - Food
Instructor(s): Mikael Kubista Course Number: 133
Affiliation: TATAA Biocenter
Course Date: 03/15/2012 - Thursday Course Length: 1 Day Course
Start Time: 08:30 AM End Time: 05:00 PM
Fee: $455 ($655 after 2/13/12) Textbook Fee:

Course Description
Please bring a laptop for class exercises. Quantitative real-time PCR (qPCR) has rapidly become the preferred technique for molecular testing of nucleic acids. From having been used mainly in the research environment it is now being introduced into diagnostics, which requires the qPCR test is standardized and the performance is controlled for quality. In this course we go through the quality aspects of qPCR, learning how to identify the weak steps in a testing process in a pilot study, how to determine sensitivity, reproducibility and limit of detection of the test, and how to design large studies based on the established performance of the test in the pilot.

Target Audience
This course is intended for experienced researchers and laboratory personnel, including project leaders, laboratory heads that perform or plan to perform qPCR studies in routine setting or larger pre-clinical or clinical studies employing qPCR.

Course Outline
• qPCR data pre-processing
• Interplate calibration for multiplate measurements
• Selecting normalization strategy
• Preparing Standard curves
• Estimating PCR efficiency and precision
• Estimating concentrations of unknowns by means of standard curves
• Determining limit of Detection
• Determining relative expression of a gene under different conditions 
• Comparing expression of multiple markers
• Setting up a pilot study
• Optimizing workflow – estimating variance contributions
• Selecting optimum markers
• Designing and cost optimizing large studies
• The MIQE guideline

Course Instructor's 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 ( 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 for Unesco Kubista introduced qPCR in Africa and in the Middle East. In 2001 Kubista founded the TATAA Biocenters (, 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 (, which is the consortium developing guidelines for pre-analytics in molecular diagnostics. The TATAA Biocenters also organize the main annual QPCR symposium ( Kubista is advisor to several of the leading companies in the qPCR field including Life Technologies and Roche.