Quantitative PCR techniques

Preliminary Course Date:  February 23-25, 2011, Breda (NL)    Last update: 15-07-2010

Aim of the course:  

Quantitative Polymerase Chain Reaction is becoming more and more important in almost all laboratories. The aim of this international short-course is to give the participants an in-depth view in the theoretical and practical knowledge of Quantitative PCR techniques. State-of-the-art experts in the field of Quantitative PCR techniques from several research and diagnostic laboratories will provide further theoretical and practical expertise.

 

Participants:

This course is developed for experienced laboratory technicians or researchers involved in the life sciences laboratories, incl. medical diagnostics, microbiology, genetics, pathology, immunology,
agriculture, biochemistry, etc.

Basic knowledge of molecular biology in general and PCR is a prerequisite. If you feel you lack the basic knowledge in PCR or the principles of real-time PCR, but you would like to participate in this quantitative PCR course, please note that the Avans Hogeschool also offers an international introductory short course: Principles, parameters, and applications of PCR (code 2545), which is especially suitable as a basic introduction to this course. 

Course structure:  
The course consists of a series of practical sessions interspersed with lectures and tutorials. 50% of the time is devoted to lectures and 50% is spent on "hands-on" experiments, covering techniques that may be directly applied in research and diagnostic laboratories. All participants will receive a practical manual containing all used protocols and a reference manual "Rapid cycle real-time PCR", Springer Verlag, Berlin.

 

Course content:

• Day 1: Pre-qPCR: Sample preparation and RNA isolation for qPCR
• Day 2: qPCR: Quantitative PCR techniques (including probelibrary)
• Day 3: Post-qPCR: Normalisation and data analysis

 

Theory:

• RNA isolation and conventional quantitative PCR techniques
• Real-time quantitative PCR and mathematics
• Trouble shooting and discussion of the qPCR problems

 

Guest Lectures:

• Quantitative and qualitative multiplex PCR: Applications in diagnosis of viral diseases
  Dr. Richard Molenkamp, Dpt. Clinical Virology, AMC, Amsterdam
• Primer and probe design for quantitative real-time PCR, Facts and puzzles
  Dr. Dario Papi, Tib-MolBiol, Berlin, Germany
• Real-time PCR: how to do successful gene expression analysis?
  Dr. Jan Hellemans, Biogazelle, Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium
• qBasePlus: Management and automated analysis of real-time qPCR
  Dr. Jan Hellemans, Biogazelle, Ghent, Belgium
• Detection of minimal residual disease with RQ-PCR
  Dr. V. van der Velden, Dept. of Immunology, Erasmus MC, Rotterdam
• Validation of array-based gene expression profiles by real-time RT-PCR.
  Dr. Sarah Forbes-Robertson, Human Cancer Studies Group, University of Swansea, Swansea, UK.
• The impact of temperature performance of real-time thermal cyclers on the generated qPCR results
  Mary Span, CYCLERtest B.V. & BIOplastics B.V.
• Development of a consistent protocol for qPCR analysis in azalea flowers
  Dr. Ellen de Keyser, ILVO-plant Unit-Applied Genetics and Breeding , Melle Belgium
  
  

Practicals:

• Pre-PCR: Sample preparation; RNA isolation of complex tissues (e.g. MagnaLyser, Polytron)
• Pre-PCR: RT-PCR (e.g. kit based RT reaction)
• Real time quantitative PCR of low copy genes; detection with SybrGreen, Probelibrary probes, and hybridization probes
• Real time quantitative PCR; Housekeeping genes which, when, and how to use them
• Post-PCR: Assumption free analysis of PCR efficiency (LinReg), genorm, qBASE, probefinder software
  

Course staff:

Course leader:
Dr. Onno Bakker, Qpcr&A, NL.

Course co-ordinator:
Dr. M.C.M. Verschuren, Avans Hogeschool, Research group Analysis Techniques in Life Sciences, Breda, NL.

 

Accreditation:

All participants will receive a certificate of attendance. This course is awarded with 36 UEC
(eurocredits for the european professional dossier), which will be mentioned on the course certificate.

 

General Information:

Code:                 2548
Registration fee:   € 1.200,-. The fee includes course materials and lunch.
Language:           English

Location:             Avans Hogeschool, Lovensdijkstraat 61, Breda, The Netherlands route description

This course can be combined with the basic course on Principles, parameters, and applications of PCR; Principles, parameters, and applications of PCR (code 2545).
Course fee of a combined registration (2545 + 2548) is € 1.600,-

 

Sponsors:  

Roche Diagnostics
Tib MolBiol
VWR International
Agilent Technologies
Stratagene
Cepheid Benelux
PrimerDesign
qBasePlus