Hands-on Application Workshops @GQ2023

Please direct your enquiry to our scientific organisation team, headed by Michael W. Pfaffl  GeneQuan.physio@ls.tum.de
 

The workshops are aimed at giving participants a deep and objective understanding of real-time quantitative PCR, digital-PCR, expression profiling, data analysis and their applications in various fields of molecular diagnostics. The courses are intended for academic or industrial researchers considering working with qPCR and/or dPCR or scientists currently working with these technologies who seek a deeper understanding.
All workshops offer extensive hands-on training by experts in the field on
Thu 23rd to 24th March 2023.

The workshop laboratories and seminar rooms are very close to the central lecture hall.
The workshops are hosted by:





In the "MicroRNA Analysis" and in the "Hands-on qPCR" workshops the qPCR data conversion, normalisation procedure, biostatistical calculations and the expression profiling will be done with the newest GenEx software by MultiD.
You can download a free trial version => Genex.Gene-Quantification.info

logo MultiD
The courses cover all aspects in quantititative PCR or digitl-PCR and take two days to complete. Each course is app. 40-50% hands-on and is limited to 20 participants, resulting in very interactive teaching where everybody is given the opportunity to try the instrumentation. After visiting the course, participants will be able to plan and perform qPCR or digital-PCR experiments themselves, as well as anlyse and interpret the generated data.
Detailed course material and full catering (lunch, coffee, soft drinks and snacks) are included in the course fee.

Course dates                23rd & 24th March 2023               
Symposium & Workshop fees
Course starts at            9 am     
Course ends around    5 pm

Please register for the Symposium & Workshops using the ConfTool registration platform => https://www.conftool.com/GQ2023/


Workshop short agendas:

Digital PCR  (2-days)  hosted by Bio-Rad
Seminar room -- S1

Description:  Digital PCR is a technology that is gaining momentum in multiple areas of nucleic acid analysis. This course introduces the user to digital PCR technology with an solid review to the theory, overview of old and new applications, hands on wet chemistry experiments (absolute quantitation, Copy Number Variation and Rare Event Detection, Genome Editing Screening for NHEJ/HDR), data analysis and math and stats related to dPCR.

Day 1 -- preliminary agenda
  • Welcome and Introductions
  • Introduction to Digital PCR
  • Experimental reaction setup
  • Abs and CNV Quant Experiment (wet)
  • ABS, CNV and RED applications overview
  • Data collection for ABS and CNV experiment
  • RED and GE experiment (wet lab)
  • ABS and CNV results analysis
  • Data collection for RED experiment
  • Review of the day
Day 2 -- preliminary agenda
  • dPCR assay design and optimization
  • ddPCR -- basic statistics
  • RED results analysis
  • Other ddPCR Applications
  • When qPCR and when ddPCR? Moving from qPCR to ddPCR
  • Open Q&A session
  • Review of the workshop


Basic hands-on qPCR  (2-days)  hosted by TATAA Biocenter
Seminar room -- S2

Target Audience
Researchers interested in analyzing nucleic acids

Experience Qualifications
Basic knowledge about DNA & RNA and molecular analyses

Description
This is a basic real-time qPCR course for people starting with qPCR or is working with the technique and want to understand it deeper. The course starts with describing how the technology works, different types of detection technologies and instruments. Then you will learn how to design primers and probes and optimize new assays. You will get an understanding of the qPCR reaction so you are able to troubleshoot and improve your own experiments. During the first day a standard curve hands-on experiment will be performed where we will practically show how to evaluate amplification curves and handle qPCR instrument software.
The second day will cover how to do relative quantification with qPCR, how to find stable reference genes and do proper normalization. Several quantification examples will be demonstrated with calculations of ΔCq, ΔΔCq and efficiency corrected ratios. During hands-on lab you will receive relative quantification data that you will perform calculations with yourself with a few different methods. You will get an understanding of how Cq-values, thresholds and efficiency affect your quantification results. During the day you will also learn how to do absolute quantification, work with standard curves and how to properly validate your qPCR assay performance.


Day 1

Introduction to PCR and qPCR
    - How does qPCR work?
    - Different detection chemistries, dyes or probes?
    - Different applications for qPCR

qPCR data evaluation
    - How does qPCR software process the data
    - How to evaluate curves and set threshold

Primer and probe design
    - How to do proper primer design
    - How to avoid primer dimer formation
    - Other important considerations for primer design
    - How to design hydrolysis probes
    - Practical exercises in primer design

Optimization of qPCR protocols
    - Which factors affect the PCR?
    - Which factors can be optimized?


Hands-on lab running qPCR analyzing unknown samples with standard curve.

Day 2

Normalization
    - Different ways to normalize
    - How to find stable reference genes

Basic quantification theory
    - Quantification methods and equations
    - How to do interplate calibration

Absolute quantification strategies
    - How to do absolute quantification
    - What is a suitable standard?

Validation of assays
    - How to determine LOD and LOQ
    - Precision estimation
    - Which controls should be used?

Quantification calculation examples 
    - Practical examples with relative quantification calculations
    - Calculations with own generated qPCR data

Hands-on lab running qPCR and calculating relative quantities with different strategies. 




MicroRNA Analysis  (2-days)  hosted by TATAA Biocenter & MultiD
Seminar room -- S3

Target Audience
Researchers interested in analyzing microRNA

Experience Qualifications
Basic knowledge about microRNA and molecular analyses

Description
This course introduces miRNA and its variant forms. Methods to extract and purify microRNA and prepare it for analysis, including quality control. Methods for microRNA expression analysis are presented and compared, to learn how to choose the best method for each study. Analysis of microRNA data is also discussed, including challenging normalization.


Day 1

    MicroRNA background including biogenesis and function
    MicroRNA extraction sampling, miRNA stability, extraction kit comparison
    Quality control of miRNA
    MicroRNA and liquid biopsies challenges of miRNA analysis in biofluids

Hands-on lab testing of quality and quantity of miRNA



Day 2

    MicroRNA quantification with microarray, nanoString, NGS and RT qPCR methods
    Comparison of miRNA quantification methods
    Experimental design and data analysis
    RT-qPCR normalization of miRNA data
    Introduction to NGS data analysis

Hands-on lab of miRNA RT-qPCR analysis including data analysis in Genex
 



Please direct your enquiry to our scientific organisation team, headed by Michael W. Pfaffl   GeneQuan.physio@ls.tum.de
  2022 - 2023