Course Information
Course Title: Real-Time PCR
Categories: 1 - Life Sciences
2 - Polymerase Chain Reaction
Instructor(s): Mikael Kubista Course Number: 34
Affiliation: TATAA Biocenter
Course Date: 03/09/2009 - Monday Course Length: 1 Day Course
Start Time: 08:30 AM End Time: 05:00 PM
Fee: $425 ($625 after 2/9/09) Textbook Fee:

Course Description
Molecular diagnostics and molecular research are experiencing a technology shift. A new platform based on ultra sensitive detection of nucleic acids - called quantitative real-time PCR (qPCR) - has been developed that makes it possible to determine the amounts of specific nucleic acids and even proteins in biological samples with unsurpassed accuracy and sensitivity. This has opened new possibilities to study and diagnose diseases based on expression patterns and to detect pathogens in biological samples. This 1-day course will give participants an in-depth understanding of real-time PCR technology and its possibilities, as well as insight in the future development of the technology platform and forthcoming applications.

Target Audience
This course is intended for researchers working or planning to work with real-time PCR, scientists with interest in gene and protein expression, clinicians and pathologists with interest in molecular diagnostics, virologists, bacteriologists, and scientists with interest in food diagnostics. Also business people and people from the financial sector interested in investing in companies active in gene expression or the diagnostic market are targeted.

Course Outline
• Basic qPCR, applications and possibilities of the technology
• Available and forthcoming instrumentation
• The most common detection methods
• Primer design and PCR optimization
• Probe design for TaqMan and Molecular Beacons
• Design and performance of qPCR experiments.
• Pre-treatment of measured data
• Strategies for normalization
• Quantification strategies
• Reverse transcription
• Extraction of DNA and RNA
• Quality assessment in qPCR
• SNP and mutation analysis using qPCR
• Multiplex qPCR
• Immune qPCR for protein detection
• Multimarker qPCR diagnostics
• Digital PCR
• Future developments in qPCR and novel applications

Course Instructor's Biography
Professor Mikael Kubista was among the pioneers who developed real-time PCR. Starting in 1991 his laboratory developed dyes and probes for real-time PCR and founded LightUp Technologies (www.lightup.se) as the first company in the world focusing on real-time PCR based human infectious disease testing. His team then developed experimental approaches for accurate measurements of expression levels by real-time PCR, and they pioneered the fields of single cell expression profiling and multidimensional expression profiling by real-time PCR. Kubista also developed methods and approaches to analyze gene expression data and founded the company MultiD Analyses (www.multid.se) that develops the popular software GenEx for real-time PCR data analysis. Working as advisor for Unesco he introduced real-time PCR in Africa and in the Middle East. In 2001 Kubista founded the TATAA Biocenters (www.tataa.com), as the world leading service providers and organizers of hands-on training in real-time PCR. Regular training courses are held all over Europe, Africa and Asia. In US TATAA Biocenter arranges courses in Santa Clara, CA, in collaboration with Intelligent Enterprise Solutions (www.iesols.com) and in collaboration with Pittcon. The TATAA courses world-wide are supported by leading instrument manufacturers and reagents suppliers in the real-time PCR field. The TATAA Biocenters organize the leading QPCR symposia in Europe (http://qpcr2007.gene-quantification.info/) as well in Europe (www.qpcrsymposium.com). Most recently Kubista set up the first high throughput qPCR expression profiling center in Europe at the new institute of Biotechnology in Prague (www.ibt.cas.cz).