Day 1:
This 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:
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, hydrolysis probes, Molecular Beacons...etc)
Different instrument platforms 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:
Introduction to quantification principles
Quantification strategies, uses and limitations
Calculations using different relative quantification methods
Strategies for normalization of qPCR data
In situ calibration for compensation of inhibition in samples
Absolute quantification
Day 3:
Sample Preparation (Extraction of RNA and DNA)
Principles of RT
Priming methods for RT
RT optimization
The MIQE guidelines
Group discussion, bring your own questions
2 days Hands-on qPCR
Day 1:
This 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:
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, hydrolysis probes, Molecular Beacons...etc)
Different instrument platforms 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:
Introduction to quantification principles
Quantification strategies, uses and limitations
Calculations using different relative quantification methods
Strategies for normalization of qPCR data
In situ calibration for compensation of inhibition in samples
Absolute quantification
3 days Experimental design and statistical data analysis for qPCR
This is a comprehensive course teaching the basics of statistics including the most common methods to analyze univariate as well as multivariate qPCR. The course includes extensive computer based exercises.
Day 1- Real-time PCR experimental design and basic data processing
Basic principles of statistics
Experimental design, selecting reference genes
Absolute quantification
Day 2- Statistical analysis of real-time PCR data
Advanced principles of statistics
Statistical tests
Ability to detect a difference
Separate variance contributions
Relative quantification, Comparison of groups
Day 3- Gene expression profiling with real-time PCR
Multiplate measurements
Normalization
Introduction to Expression Profiling
Data preprocessing
Classification methods
2 days Experimental design and statistical data analysis for qPCR
The two-days course covers the most of the same prinicples as the three-day analysis course but less comprehensive and with less practical excercises
Day 1- Statistical analysis of real-time PCR data
Basic principles of statistics
Advanced principles of statistics
Statistical tests
Ability to detect a difference
Day 2- Gene expression profiling with real-time PCR
Multiplate measurements
Standard curves and absolute quantification
Experimental design, Selecting reference genes
Relative quantification, Comparison of groups
Expression profiling
2 days Single cell analysis
In this course you will learn to recognize the difference
between cell population and single-cell analysis and how to interpret and apply
single-cell data in biological studies. Practical training in cell collection,
gene expression profiling and data analysis at the single-cell level is included. The course covers:
Overview of single-cell biology: methods,
examples and trends
Cell population versus single-cell
analysis using gene expression profiling
Cell collection methods. Demonstration
of using FACS to collect single-cells
Practical exercise running RT-qPCR to
analyze individual cells
Single-cell data analysis, including
practical exercises
1 day Sample preparation and quality control of nucleic acids
One of the most important requirements to get good results from qPCR experiments 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. It also includes the important quality control steps needed to asure reliable results. The course covers:
Overview of nucleic acid extraction methods
How to properly determine the concentration of purified nucleic acids
Storage of biological samples and purified nucleic acids
Quality control of purified material
Integrity of purified RNA
How to test for the presence of inhibitors.
1 day High throughput and digital PCR
Lectures cover the design of large expression profiling experiments, including plate design and interplate calibration as well as the pre-processing of gene expression profiling data. During the course you will also learn how to run digital PCR for precise quantification using chip-based instrument. Hands-on experiments will be performed on chip-based high throughput instrument. More info will be added soon.
1 day Immuno-qPCR
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
A technology description
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
1 day Multiplex PCR
This is a 1-day course in multiplex PCR with a focus on multiplex real-time PCR. The course consists of a theoretical part and a practical part where participants get to do multiplex qPCR experiments by themselves under experienced supervision. The course contains:
Introduction to multiplex PCR
Multiplex PCR applications
Primer design
Instrument, filter set and dye considerations
Optimization of multiplex assays
Validation of multiplex assays
1 day HRM
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
Gene scanning
Methylation analysis
Review of available HRM instruments
Examples of other applications
1 day qPCR for microRNA analysis
This is an introductory course to miRNA analysis using qPCR. The course consists of a theoretical part and a
practical part where participants get to do miRNA experiments
by themselves under experienced supervision. The course covers:
Introduction to miRNA
Applications, what can miRNA analysis be used for?
How to extract and isolate miRNA
Different ways to quantify miRNA with qPCR
Normalization
Data analysis and interpretation
1 day Quality control of qPCR in molecular diagnostics
This course will give you the tools in how to do proper quality control of your qPCR assays to be used in molecular diagnostics. It will describe which controls that is needed, how they should be performed and the statistics on how to do the evaluations.
