All four
workshops are hosted by BioEPS GmbH (www.bioeps.com) andTATAA
Biocenter
Germany (http://workshops.gene-quantification.info) The qPCR
workshop laboratories and seminar rooms are very close to
the central lecture hall.
The workshops are
aimed at giving participants a deep and objective understanding of
real-time quantitative PCR, biostatistics, expression profiling, and
its applications. The courses are
intended for academic or industrial persons considering working with
qPCR or scientists
currently working with qPCR seeking a deeper understanding. Please
have a
look to the workshops of the next
workshops
Four different
2-day workshops will be held in parallel at 12th to 13th March
at the qPCR 2009 Event:
All
workshops offer extensive hands-on training by qPCR experts from BioEPS GmbH team (www.bioeps.com) Leading probe technologies will be
demonstrated,
common and emerging instrument platforms will be available and advanced
primer and probe design programs will be used. Hands-on training will
be provided on quantitative gene expression, including design and
optimization of both RT and qPCR protocols.
Participating companies are asked to support the workshops with PCR
cyclers, qPCR kits, consumables, centrifuges, pipettes etc…. Please
contact Michael Pfaffl or Martina Reiter at bioEPS GmbH or the TATAA
Biocenter Germany Martina.Reiter@bioEPS.com
for information and details.
The
course covers all aspects in qPCR and is held during 2-days. Each
course is app. 50%
hands-on and is limited to app. 30
participants,
resulting in very interactive teaching and everybody given the
opportunity to try the instrumentation.
After
the course participants will be able to plan and perform qPCR
experiments themselves, as well as interpret and analyze data. Detailed
course material and full catering (lunch, coffee, soft drinks and
snacks) are included in the course fee.
The
introductory course consists of a theoretical part and a practical part
where participants get to do QPCR experiments by themselves under
experienced supervision. The course contains:
Day 1:
•
Basic PCR theory
• The theory of real-time PCR
• Applications and possibilities of QPCR. Comparison
of QPCR with regular PCR.
• Review of currently available detection
technologies (SYBR Green I, TaqMan, Molecular Beacons...etc)
• Different instrument plattforms and their typical
uses
• Primer Design
• The problem of primer-dimer formation and how to
minimize them
• Probe Design
• Experimental design and optimization
• Basic data handling and analysis
Day 2:
This course
covers aspects in sample preparation and reverse transcriptions.
• Principles of RT
• Priming methods for RT
• What enzymes are preferential for different
applications
• Sample Preparation (Extraction of RNA and DNA)
• Introduction to statsitics and statistical analysis
of data
This course explains
statistics applicable to qPCR and teaches how to use statistics to
interpret real-time PCR gene expression data, and classify samples
based on real-time PCR expression profiling. Course is based on
seminars
and computer-based demonstrations. Please
bring your own Laptop
to the course!
Day
1- Statistical analysis of real-time PCR data Lectures cover the principles of statistics, including
Gaussian statistics, the central limit theorem, p values and
statistical hypothesis testing, z-scores, rank-based methods
(non-Gaussian), comparison of two groups (paired and unpaired t-test),
Mann Whitney test, Wilcoxon rank sum test, Fisher’s exact test. Outlier
detection (Dixon’s test, Grubb’s test, Cochran’s test), ANOVA and
classical calibration (least square fit, correlation coefficient,
Hottelings’ area). During computer based workshop participants will
learn how to analyze typical real-time PCR data sets. Examples include
identification of outliers, and how to compare means and variances of
paired and unpaired studies.
Day
2- Gene expression profiling with real-time PCR Lectures cover methods to classify samples and genes.
The methods presented include Principal Component Analysis, Potential
Curves, Hierachical Clustering, Self-Organizing Maps, and Trilinear
Decomposition. During computer based workshops participants will
classify metabolic genes in yeast, developmental stages in Xenopus
laevis, Breast cancer data, and developing stem cells.
Sample
Preparation
(2-days)
One
of the most important requirements to get good results from qPCR
experiment is to have a template of good quality. In most cases this
means having an efficient sample preparation. This course module is
focused on extraction of RNA and DNA to be used as template in qPCR and
reverse transcription reactions. The course covers:
• Overview of nucleic acid extraction methods
• How to properly determine the concentration of
purified nucleic acids
• Extraction from limited amount of material and
single cells
• Storage biological samples and purified nucleic
acids
• Quality control of purified material
• Integrity of purified RNA
• How to test for the presence of inhibitors
HRM (on day 1)
& Immuno-qPCR (on
day 2):
High-Resolution_melting
(HRM on day 1)
This
is an introductory course in HRM, where a high resolved melting curve
is used to analyze very small differences in melting temperature of PCR
products, differences that can be due to a single base substitution.
The course includes seminars as well as hands on training where the
participants get to perform experiments. The course covers:
•
Introduction to HRM
• Assay design
• SNP – analysis
• Genescanning
• Methylation analysis
• Review of available HRM instruments
• Examples of other applications
Immuno-qPCR
(on day 2)
This
course shows how real-time PCR can be used to quantify proteins. The
course consists of a theoretical part which explains what immuno-qPCR
is and how it can be set up and used. The course also includes a
practical part where the course participants will run an immuno-qPCR
experiment to quantify a protein. The course covers:
•
Introduction to immunoassays. •
How to set up an immuno-qPCR assay. •
How to optimize an immuno-qPCR. •
How to analyse immuno-qPCR data. •
Troubleshooting. •
Examples of immuno-qPCR applications. •
Practical experiment quantifying a protein.