Organized by

Logo-INRAE_Transparent.png
  • Facebook
  • Twitter
  • YouTube

MICROGENOMICS Technical Workshop

 September 9, 2020 at the University of Florence, Italy

Plenary session with an Introduction to Microgenomics and time dedicated to Gold and Silver Sponsor presentations 

 

4 one-day technical workshops (simultaneous) organized by an academic scientist


    1- Methods and strategies for molecular biomarker analyses

         Scientific leader : Mikael Kubista, Prof.


    2- Rare cell analysis : from enrichment to single cell analysis

          Scientific leader : Pamela Pinzani, Prof.


    3- EV relevant guidelines, EV & Exosome Isolation, and on the EV & Exosome

        characterisation

        Scientific leader : Michael W. Pfaffl, Prof.


    4- Digital PCR and Microgenomics (soon to be available)

 > Sponsor stands during lunch and cocktail with sponsors.

Preliminary agenda

1- Methods and strategies for molecular biomarker analyses
Scientific leader : Mikael Kubista, Prof.

Target audience: Researchers and clinicians at all levels interested in molecular methods for biomarker analyses 

Entrance qualifications: Basic knowledge of molecular methods 

Description: This course will present methods for nucleic analysis covering SNP genotyping, rare mutation analysis, methylation analysis, copy number analysis, gene expression analysis, splice variants, microRNA and exosome analysis. The whole workflow from sampling, preanalytics, analytics and data analysis, and relevant guidelines are discussed, The course is lecture based.

 

The course contains:

  • Introduction to molecular analysis methods

  • Methods for SNP analysis incl. qPCR, HRM and sequencing

  • Introduction to rare mutation detection, copy number variation and variance analyses

  • Methods for rare mutation analysis incl. CastPCR, RNase H dependent PCR, myT primers, Two-Tailed PCR, dPCR, SimSen Seq etc.

  • Methods for methylation analysis and other epigenetic variations.

  • Introduction to gene expression analysis 

  • Methods for expression analysis incl. qPCR and sequencing

  • Introduction to microRNA and exosome analyses

  • Methods for microRNA analysis including microarrays, sequencing, qPCR and Two-Tailed PCR

  • New guidelines and regulations for molecular analyses including CEN, ISO and IVD-R

  • Quality control procedures in molecular analyses.

2- Rare cell analysis: from enrichment to single cell analysis
Scientific leader : Pamela Pinzani, Prof.

The analysis of rare cells in complex and heterogeneous samples with a large population of background cells has gained a relevant role in different fields such as cancer diagnosis and prognosis, prenatal diagnosis, viral infections, forensic applications, etc.

 

The most studied rare cells are represented by circulating tumor cells in the blood of cancer patients defined as a “liquid biopsy” of the tumor, but also other populations of cells, such as tumor stem cells, endothelial cells, hematopoietic progenitor cells and their subpopulations, fetal cells in maternal blood, have gained importance in biomedical research.

 

The analysis of rare cells is challenging and requires complex multi-step processes starting from sample collection and ending with single cell molecular analyses.

New technologies are continuously emerging and evolving in this field and there is a need of integrating different methods in standardized and reproducible workflows to translate rare cell analysis into clinical practice.

 

The aim of the workshop is to give an overview of up to date principles and methods for rare cell analysis, even at the single cell level.

Critical aspects of the workflow for the analysis of rare cells will be addressed and solutions proposed.

A focus on particular technologies will be provided.

 

Tentative program:

  • Introduction to rare cell technologies

  • Rare cell enrichment by immunomagnetic capture

  • Rare cell enrichment on the basis of cell physical properties

  • Rare cell enrichment/isolation by flow cytometry

  • Single Cell sorting: dielectrophoresis, micromanipulation, laser-microdissection, etc.

3- The EV workshop focus will be on EV relevant guidelines, EV & Exosome Isolation, and on the EV & Exosome characterisation.
Scientific leader : Michael W. Pfaffl, Prof.

1) How to standardize the EV workflow:
    Talk on MISEV guidelines and EV track 
2) EV Isolation:
    Practical workshop on different isolation methods, the underlying principles (company

    presentations)
3) EV characterisation:
    How to characterize EVs according to the MISEV guidelines
    NTA:  absolute size ans size distribution (company presentation and talk)
    TEM:  electron microscopy (invited talk)
    Western Blotting:  EV surface protein marker (company talk)
    NGS:  small-RNA Seq and data analysis (invited talk)
    Proteomics:  LC-MS (invited talk)
4) Round table & Final Discussion