Real time Polymerase Chain Reaction

Real time Polymerase Chain Reaction is becoming more and more important in almost all laboratories. Especially in diagnostic laboratories real time PCR is a new, and promising, technique to identify bacteria, viruses and genetic alterations in human cells. The aim of this international short-course is to give the participants an in-depth view in the theoretical and practical knowledge of real time PCR techniques in molecular diagnostics. State-of-the-art experts in the field of real time PCR techniques from several diagnostic laboratories will provide theoretical and practical expertise.
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This course is developed for experienced laboratory technicians involved in the life sciences laboratories, incl. medical diagnostics, microbiology, genetics, pathology, immunology, 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 basic 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 diagnostic laboratories. All participants will receive a practical manual containing all used protocols.

Course content:

Day 1: Pre-PCR: Sample preparation and RNA isolation for real-time PCR and HRMA
Day 2: Real-time PCR and validation
Day 3: HRMA, digital PCR and Post-qPCR data analysis

Theory:

Sample preparation for real-time PCR
RNA quality control (Lab-on-a-chip, RIN)
Real-time quantitative PCR and mathematics
Trouble shooting and discussion of qPCR problems

list of Guest Lectures:

Primer and probe design for real time PCR in molecular diagnostics, Facts and puzzles
Dr. Dario Papi, Tib-MolBiol, Berlin, Germany
Quantitative and qualitative multiplex PCR: Applications in diagnosis of viral diseases
Dr. Richard Molenkamp, Dpt. Clinical Virology, AMC, Amsterdam
Detection of minimal residual disease with RQ-PCR
Dr. V. van der Velden, Dept. of Immunology, Erasmus MC, Rotterdam
How to deal with qPCR thermocycler variability in moleculair diagnostics
Mary Span, CYCLERtest B.V. & BIOplastics B.V.
High resolution melting curve analysis in molecular diagnostics.
Leendert Vermeulen, Roche Diagnostics, Almere
Inreoduction to droplet digital PCR, overview and applications of the 3rd generation PCR
E. van Collenburg, Bio-Rad laboratories, Veenendaal
Practicals:

Pre-PCR: Sample preparation; RNA isolation from virus, and leukemia cells
Real-time PCR: Hybridization probe analysis: HSV1 versus HSV2
Real-time PCR: Multiplex real-time PCR for Respiratory viruses
Real time quantitative PCR: Real time quantitative PCR for BCR-ABL translocations (MRD)
Real-time PCR: High resolution melting curve analysis: HSV1 versus HSV2
Post-PCR: qPCR data analysis of real-time quantitative PCR data

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.

24 Oct - 26 Oct 2012

Breda
Netherlands
meeting website