qPCR course

Important note!

In 2016, the Doctoral Schools will not refund the qPCR course followed at Biogazelle.

Registration fee - Cancellation policy

This course is free of charge for members of the Doctoral School of Life Sciences and Medicine, (Bioscience) Engineering and Natural Sciences. A no show fee of 250 EUR will be charged for cancellations without valid reason after October 5th, 2017. This policy is applied due to the high cost of the course and the number of interested students on the waiting list.

Target audience

The course is targeted to doctoral students belonging to all Science Doctoral Schools of Life Sciences and Medicine, Natural Sciences and Bioscience Engineering. Students from all three Doctoral Schools are potentially challenged with qPCR experiments during their doctoral research. This course is intended to train and to create awareness about good practises in the performance of qPCR experiments amongst researchers from different disciplines. Morever, the course is fully aligned with the MIQE guidelines (Bustin et al., 2009), encouraging better experimental practice, more reliable and inequivocal interpretation of qPCR results.
The course will be of benefit not only for the attendees but also for the co-workers of the laboratories in which the attendees perform their research, as the course could serve to set standards for a proper in-house qPCR workflow.

Topic and theme

During the last decades, the number of researchers from different disciplines performing transcriptome analysis has increased dramatically. Despite the use of state-of-the-art highthroughput technologies for whole genome transcriptome profiling (e.g. next generation sequencing or microarray), quantitative real-time PCR (qPCR) remains the gold standard for targeted gene expression analysis due to its superior sensitivity, specificity, linear dynamic range of quantification and a high level of flexibility. A qPCR experiment involves several steps ranging from experimental design, RNA extraction, assay design, data generation to data analysis. While the practical performance of a real-time PCR quantification experiment is relatively straightforward, it is important to identify critical steps that may impact the quality and robustness of the experiment. Moreover, many researchers experience a genuine need for more in-depth training of their experiment design and dataanalysis
skills in order to generate biologically relevant and reliable results. To accommodate these needs, Biogazelle offers a qPCR-focused course in which the fundamental and advanced principles of experiment design, sample and assay quality control and dataanalysis are covered.

Lecturers

dr. Jan Hellemans, CEO Biogazelle has a Master in Biotechnology (2000, UGent) and a PhD in Medical Genetics (2007, UGent). He is author of more than 100 scientific articles in international journals, including some pioneering publications in the domain of real-time PCR. Together with professor Jo Vandesompele, he is active in the development of new concepts and algorithms for improved qPCR data analysis, and one of the founders of the RDML consortium (globally accepted qPCR communication protocol). Jan Hellemans supervised a core facility for automated DNA-isolation, (next generation) sequencing and high-throughput real-time PCR at Ghent University. He is a co-founder and CEO of Biogazelle, active as a teacher in qPCR courses and responsible for dry lab activities at Biogazelle.


dr. Bram De Craene, Post-doctoral researcher VIB, Training and support Biogazelle, has a Master in Biotechnology (2001, UGent) and a PhD in Sciences (2005, UGent). He worked as postdoctoral researcher at the Inflammation Research Center department of VIB, where he was managing the departamental qPCR facility, and is author of several scientific articles in the cancer research field. Currently, Bram works as Senior Scientist assay design and development at Biocartis. Moreover, he works as trainer for Biogazelle’s qPCR courses and in the technical support for the qPCR data analysis software qbase+.

Programme and course content

The course comprises different chapters covering the whole qPCR procedure from experimental design to data generation and interpretation of the results. Practical examples and exercises are given for each section, allowing participants to interact and formulate their questions. For each chapter, a balanced mix between theoretical background and practical hands-on data analysis is included. During the course, practical examples using the qPCR data analysis software qbase+ are foreseen and a 2-months Premium qbase+ license is provided to each attendee.

  • 1. Experiment design

• Power analysis
• Experiment layout (sample vs gene maximization)
• Biological and technical replicates
• Inter-run calibration

  • 2. Sample preparation and quality control

• RNA integrity and purity
• DNase treatment
• cDNA synthesis
• Pre-amplification

  • 3. Assay design and validation

• Primer design
• Probes vs intercalating dye
• Design guidelines
• In silico evaluation
• Empirical validation

  • 4. qPCR amplification

• Real-time PCR principle
• Amplification curve
• Melt curve
• Cq value determination methods
• Replicates and controls
• Speed and throughput considerations

  • 5. Absolute quantification

• Standard curves
• Limitations and concerns

  • 6. Relative quantification

• Quantification models
• Efficiency correction
• Selection and validation of reference genes
• Normalization with multiple reference genes
• Alternative normalization methods
• Inter-run calibration
• Result rescaling

  • 7. Data quality and control

• Visual inspection of plots
• Positive and negative controls
• Normalization factor histogram
• Reference gene stability
• Inter-run variation

  • 8. Bio-statistical analysis

• Basic principles
• Descriptive statistics
• Selection and application of appropriate statistical tests

Dates and venue

  • Thursday 19 + Friday 20 October 2017 from 9:00 - 17:00 (1h lunch break from 12:30 till 13:30)

Campus Coupure, Coupure links 653, 9000 Gent

Building B, ground floor, room 0B.037

  • Important: bring your own laptop to install the software!

Registration

Please follow this link: https://webapps.ugent.be/eventManager/events/qpcrbiogazelle  Your registration will be confirmed by e-mail.

When the course is fully booked, please ask by e-mail to   to be added to the waiting list which means a seat will be reserved for you during the next course. Please note that maximum 20 participants are allowed.

Evaluation criteria (doctoral training programme)

100% participation

Course material

• One week before the course, an email will be sent to all attendees providing them with an activation code and instructions to install qbase+
• On the first day of the course, a hard copy from the presentations will be delivered
• Activation code valid for a 2-months qbase+ license
• Certificate of attendance

Number of participants

Maximum 20