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Gene Quantification
Q2 / 2018

is sponsored by

    Dear researcher,
dear Gene Quantification page reader,

Our newsletter informs about the latest news in gene expression profiling using qPCR, dPCR, NGS, and related methods, which are compiled and summarised on www.Gene-Quantification.info

The focus of this qPCR NEWS issue is:




Streaming Server


Join the updated
eConferences streaming webportal
www.eConferences.de -- Amplify your knowledge in  qPCR, dPCR and NGS !
Free access to over 350 recorded talks. This streaming portal is dedicated to scientists from the community of qPCR, digital PCR, Next Generation Sequencing (NGS), MicroGenomics (MG) and Molecular Diagnostics (MDx). You’ll find here all the records of the presentations held at qPCR dPCR & NGS and MG Events in the past years qPCR 2010 in Vienna till qPCR dPCR & NGS 2017 in Freising-Weihenstephan.
We provide the presentations via movie streaming technology in high quality – high resolution and perfect sound quality in high speed – on any internet browser or mobile device.


Extracellular Vesicles (EV) and Exosomes in Molecular Diagnostics

Extracellular Vesicles (EVs) and exosomes are cell-derived vesicles that are present in many tissues and (perhaps) in all biological fluids, including blood, urine, milk, cerebrospinal fluid, saliva, sweat and also in any kind of cell culture supernatant. The reported size of exosomes is between 50 and 200 nm in diameter. EVs are bigger and range between 150 up to 500 nm. It is becoming increasingly clear that especially exosomes have specialized functions and play a key role in inter-cellular communication, e.g. in the immune system, in cancer progression, in coagulation, in inter-cellular signaling, in cellular waste management, and even it is assumed that they are important in the regulation of any physiological process.
Consequently, there is a growing interest in molecular diagnostic and in clinical application of EVs with a focus on exosomes. EVs and exosomes can potentially be used for prognosis, therapy, and biomarkers for health or disease, most applied today in cancer diagnostics and progression.
A lot of research is done in EV and exosome purification, isolation, size determination, and content characterization. They contain various molecular constituents of their cell of origin, including cell-surface & intra-cellular proteins, membrane fatty acids, and various types of RNAs. Especially the high concentrated regulative small-RNAs, e.g. the microRNAs, are of major interest.

EV-TRACK  --  transparent reporting and centralizing knowledge in extracellular vesicle research.
EV-TRACK Consortium, Van Deun J, Mestdagh P, Agostinis P, ... and much more, Vandesompele J, Hendrix A
Nat ureMethods. 2017 14(3): 228-232

We argue that the field of extracellular vesicle (EV) biology needs more transparent reporting to facilitate interpretation and replication of experiments. To achieve this, we describe EV-TRACK, a crowdsourcing knowledgebase (http://evtrack.org) that centralizes EV biology and methodology with the goal of stimulating authors, reviewers, editors and funders to put experimental guidelines into practice.

Selected Papers & Reviews  

The exosomal RNA content in EVs and Exosomes

Obstacles and opportunities in the functional analysis of extracellular vesicle RNA - an ISEV position paper.
Mateescu B, Kowal EJ, van Balkom BW, Bartel S, Bhattacharyya SN, Buzás EI, Buck AH, de Candia P, Chow FW, Das S, Driedonks TA, Fernández-Messina L, Haderk F, Hill AF, Jones JC, Van Keuren-Jensen KR, Lai CP, Lässer C, Liegro ID, Lunavat TR, Lorenowicz MJ, Maas SL, Mäger I, Mittelbrunn M, Momma S, Mukherjee K, Nawaz M, Pegtel DM, Pfaffl MW, Schiffelers RM, Tahara H, Théry C, Tosar JP, Wauben MH, Witwer KW, Nolte-'t Hoen EN
J Extracell Vesicles. 2017 6(1): 1286095 -- eCollection 2017

Summary of the ISEV workshop on extracellular vesicles as disease biomarkers; Meeting Report -- held in Birmingham, UK, during December 2017
Aled Clayton, Dominik Buschmann, J. Brian Byrd, David R. F. Carter, Lesley Cheng, ....................., Kenneth W. Witwer, Hang Yin & Rienk Nieuwland
Journal of Extracellular Vesicles 2018 (7)1

Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next-generation sequencing.
Buschmann D, Kirchner B, Hermann S, Märte M, Wurmser C, Brandes F, Kotschote S, Bonin M, Steinlein OK, Pfaffl MW, Schelling G, Reithmair M
J Extracell Vesicles. 2018 7(1): 1481321 -- eCollection 2018

The impact of disparate isolation methods for extracellular vesicles on downstream RNA profiling.
Van Deun J, Mestdagh P, Sormunen R, Cocquyt V, Vermaelen K, Vandesompele J, Bracke M, De Wever O, Hendrix A.
J Extracell Vesicles. 2014 Sep 18;3 -- eCollection 2014.

A comprehensive method for identification of suitable reference genes in extracellular vesicles.
Gouin K, Peck K, Antes T, Johnson JL, Li C, Vaturi SD, Middleton R, de Couto G, Walravens AS, Rodriguez-Borlado L, Smith RR, Marbán L, Marbán E, Ibrahim AG
J Extracell Vesicles. 2017 6(1): 1347019 -- eCollection 2017

Exosomes provide a protective and enriched source of miRNA for biomarker profiling compared to intracellular and cell-free blood.
Cheng L, Sharples RA, Scicluna BJ, Hill AF
J Extracell Vesicles. 2014 Mar 26;3 -- eCollection 2014

Cellular and extracellular miRNAs are blood-compartment-specific diagnostic targets in sepsis.
Reithmair M, Buschmann D, Märte M, Kirchner B, Hagl D, Kaufmann I, Pfob M, Chouker A, Steinlein OK, Pfaffl MW, Schelling G
J Cell Mol Med. 2017 21(10): 2403-2411

The microRNA spectrum in 12 body fluids.
Weber JA, Baxter DH, Zhang S, Huang DY, Huang KH, Lee MJ, Galas DJ, Wang K.
Clin Chem. 2010 56(11): 1733-1741

Toward reliable biomarker signatures in the age of liquid biopsies - how to standardize the small RNA-Seq workflow.
Buschmann D, Haberberger A, Kirchner B, Spornraft M, Riedmaier I, Schelling G, Pfaffl MW
Nucleic Acids Res. 2016 44(13): 5995-6018



First announcement

9th Gene Quantification Event


qPCR dPCR & NGS 2019
9th international Gene Quantification Event
Symposium  &  Industrial Exhibition  &  Application Workshops
18 - 22 March 2019,  in Freising-Weihenstephan, School of Life Sciences,
Technical University of Munich, Weihenstephan, Germany


GenEx 6.1

The most powerful tool for complex qPCR data analysis


GenEx offers advanced methods to analyze real-time qPCR data with simple mouse clicks

GenEx is the most popular software for qPCR data processing and analysis. Built in a modular fashion GenEx provides a multitude of functionalities for the qPCR community, ranging from basic data editing and management to advanced cutting-edge data analysis. GenEx 6.1 – the software compliant with MIQE and CLSI guidelines.

TATAA Biocenter, Europe´s leading provider of genomic services using quantitative real-time PCR (qPCR), and MultiD Analyses, Europe’s prime software developer for the analysis of multivariate data, release GenEx version 6 for accurate qPCR data analysi, compliant with current MIQE guideline and Clinical Laboratory Standards Institute (CLSI) guideline.

GenEx 6.1

international GenEx software presentations


Listen to GenEx presentations:

GenEx 6.1

Compliant with MIQE and CLSI guidelines


Basic data editing and management
Arguably the most important part of qPCR experiments is to pre-process the raw data into shape for subsequent statistical analyses. The pre-processing steps need to be performed consistently in correct order and with confidence. GenEx Standard’s streamlined and user-friendly interface ensures mistake-free data handling. Intuitive and powerful presentation tools allow professional illustrations of even the most complex experimental designs.

Advanced cutting-edge data analysis
When you need more advanced analyses GenEx Enterprise is the product for you. Powerful enough to demonstrate feasibility it often proves sufficient for most users demands. Current features include parametric and non-parametric statistical tests, Hierarchical Cluster Analysis (HCA), Heatmap, Principal Component Analysis (PCA, and Artificial Neural Networks. New features are continuously added to GenEx with close attention to customers’ needs.

New features
Sample handling and samples individual biology often contribute to confounding experimental variability. By using the new nested ANOVA feature in GenEx version 5 user will be able to evaluate variance contributions from each step in the experimental procedure. With a good knowledge of the variance contributions, an appropriate distribution of experimental replicates can be selected to minimize confounding variance and maximize the power of the experimental design! For experiments with complex features, such as for example multifactorial diseases, analytical relationships and classifications may not readily be available. The support vector machine feature in the new version of GenEx is so easy to use that it will make this advanced supervised classification method easily available to novice users, while providing access to advanced parameters for experts.

View our webpage and download a FREE GenEx 6.1 trial version => GenEx.gene-quantification.info



Best regards,

Michael W. Pfaffl
responsible Editor of the Gene Quantification Pages



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The qPCR NEWS and the Gene Quantification Pages are educational sites with the only purpose of facilitating access to qPCR related information on the internet.  The qPCR NEWS and the Gene Quantification Pages are edited by Michael W. Pfaffl.   Copyright ©2004-2018    All rights reserved.   Any unauthorized use, reproduction, or transfer of this or its contents, in any medium, is strictly prohibited. Disclaimer & Copyrights are displayed on our homepage  www.gene-quantification.info
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