qPCR NEWS - July 2012 - focus on microRNA
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Dear researcher,
dear Gene Quantification page reader,

Our newsletter informs about the latest news in quantitative real-time PCR 
(qPCR and RT-qPCR), which are compiled and summarised on the 
Gene Quantification homepage. The focus of this newsletter issue is:

* Update on integrated analysis of microRNA and mRNA expression - 
http://integrated-analysis.Gene-Quantification.info
* Update of new microRNA reviews and interesting microRNA normalisation papers 
- http://microRNA.Gene-Quantification.info
* Second announcement qPCR & NGS Symposium in March 2013 - 
http://www.qPCR-NGS-2013.net 
* GenEx - a powerful tool For qPCR data analysis - download a free trial 
version  -  http://GenEx.gene-quantification.info



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Quantification of miRNA-mRNA interactions
Muniategui A, Nogales-Cadenas R, Vázquez M, L Aranguren X, Agirre X, Luttun A, 
Prosper F, Pascual-Montano A, Rubio A.
Group of Bioinformatics, CEIT and TECNUN, University of Navarra, San Sebastian, 
Spain.
PLoS One. 2012;7(2):e30766. Epub 2012 Feb 14.
There is also a web-based tool for human miRNAs at http://talasso.cnb.csic.es

miRNAs are small RNA molecules (' 22nt) that interact with their corresponding 
target mRNAs inhibiting the translation of the mRNA into proteins and cleaving 
the target mRNA. This second effect diminishes the overall expression of the 
target mRNA. Several miRNA-mRNA relationship databases have been deployed, most 
of them based on sequence complementarities. However, the number of false 
positives in these databases is large and they do not overlap completely. 
Recently, it has been proposed to combine expression measurement from both 
miRNA and mRNA and sequence based predictions to achieve more accurate 
relationships. In our work, we use LASSO regression with non-positive 
constraints to integrate both sources of information. LASSO enforces the 
sparseness of the solution and the non-positive constraints restrict the search 
of miRNA targets to those with down-regulation effects on the mRNA expression. 
We named this method TaLasso (miRNA-Target LASSO).We used TaLasso on two public 
datasets that have paired expression levels of human miRNAs and mRNAs. The top 
ranked interactions recovered by TaLasso are especially enriched (more than 
using any other algorithm) in experimentally validated targets. The functions 
of the genes with mRNA transcripts in the top-ranked interactions are 
meaningful. This is not the case using other algorithms.TaLasso is available as 
Matlab or R code. There is also a web-based tool for human miRNAs at 
http://talasso.cnb.csic.es


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Joint analysis of miRNA and mRNA expression data
Muniategui A, Pey J, Planes F, Rubio A.
Brief Bioinform. 2012 Jun 12.

miRNAs are small RNA molecules ('22 nt) that interact with their target mRNAs 
inhibiting translation or/and cleavaging the target mRNA. This interaction is 
guided by sequence complentarity and results in the reduction of mRNA and/or 
protein levels. miRNAs are involved in key biological processes and different 
diseases. Therefore, deciphering miRNA targets is crucial for diagnostics and 
therapeutics. However, miRNA regulatory mechanisms are complex and there is 
still no high-throughput and low-cost miRNA target screening technique. In 
recent years, several computational methods based on sequence complementarity 
of the miRNA and the mRNAs have been developed. However, the predicted 
interactions using these computational methods are inconsistent and the 
expected false positive rates are still large. Recently, it has been proposed 
to use the expression values of miRNAs and mRNAs (and/or proteins) to refine 
the results of sequence-based putative targets for a particular experiment. 
These methods have shown to be effective identifying the most prominent 
interactions from the databases of putative targets. Here, we review these 
methods that combine both expression and sequence-based putative targets to 
predict miRNA targets.

More papers about the integrated analysis of microRNA and mRNA expression => 
http://integrated-analysis.Gene-Quantification.info


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New microRNA reviews
Technology features, pitfalls and recommendations for microRNA expression 
profiling
http://microRNA.Gene-Quantification.info

- MicroRNA profiling: approaches and considerations
- The widespread regulation of microRNA biogenesis, function and decay
- Strengths and limitations of laboratory procedures for microRNA detection
- Editorial - Studying microRNAs in the brain: technical lessons learned from 
the first ten years
- Pitfalls and recommendations for microRNA expression analysis using qPCR
- Potential pitfalls in microRNA profiling
- Technolgy Feature - MicroRNA profiling: separating signal from noise
- How Do MicroRNAs Regulate Gene Expression?
- Normalization strategies for microRNA profiling experiments: a 'normal' way 
to a hidden layer of complexity?
- The microcosmos of cancer
- miRNAs in human cancer


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New interesting microRNA normalisation papers
http://microRNA.Gene-Quantification.info

- Overview and workflow in microRNA normalisation
- Whole-Genome RT-qPCR MicroRNA Expression Profiling
- Profound Effect of Profiling Platform and Normalization Strategy on Detection 
of Differentially Expressed MicroRNAs - A Comparative Study
- Identification of reference microRNAs and suitability of archived hemopoietic 
samples for robust microRNA expression profiling
- Identification of suitable reference genes for qPCR analysis of serum 
microRNA in gastric cancer patients
- Suitable reference genes for relative quantification of miRNA expression in 
prostate cancer
- Comprehensive human adipose tissue mRNA and microRNA endogenous control 
selection for quantitative real-time-PCR normalization
- MicroRNA expression profiling to identify and validate reference genes for 
relative quantification in colorectal cancer
- miRNA expression profiling - from reference genes to global mean normalization
- Identification by Real-time PCR of 13 mature microRNAs differentially 
expressed in colorectal cancer and non-tumoral tissues
- Expression profiling of microRNA using real-time quantitative PCR, how to use 
it and what is available
- A novel and universal method for microRNA RT-qPCR data normalization


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Second announcement qPCR & NGS Symposium in Freising-Weihenstephan
18-22 March 2013
http://www.qPCR-NGS-2013.net
Download the latest Event Flyer => http://tinyurl.com/qPCR-NGS-2013

On behalf of the Organisation Committee and the Scientific Board it is a great 
pleasure to invite you to the 6th International qPCR & NGS 2013 Event. The 
event is divided in a 3-day scientific Symposium with an Industrial Exhibition 
and various 2-day Application Workshop to be held at the Center of Life Science 
in Freising Weihenstephan, Technische Universität München (Germany). The great 
international interest in the previous meetings ( qPCR 2004 to qPCR 2011 ) led 
us to the decision to repeat the Symposium in spring 2013. We expect 500-600 
participants coming from all over the world, in 2011 we could welcome 
participants from 56 contries, and roughly 30-40 international companies in the 
qPCR Industrial Exhibition.

We have set the date for the qPCR & NGS 2013 Event to 18th - 22nd March 2013. 
The event location is the central lecture hall complex and the foyer at TUM 
(Technical University of Munich) in Freising Weihenstephan, Germany (Google 
Maps link or Google Earth link). The TUM and the Biotech region around Munich 
is part of the largest Biotech cluster in Europe, located close to the Munich 
airport in the heart of Bavaria.

The focus of the qPCR & NGS 2013 Event will be on:   Next Generation Thinking 
in Molecular Diagnostics

Leading academic researchers and industrial contributors in the field will 
participate in the symposium, which will be an arena for fruitful discussions 
between researchers of different backgrounds. The Symposium Talks, Poster 
Sessions, Industrial Exhibition and associated qPCR & NGS Application Workshops 
offer an overview of the present knowledge and future developments in qPCR, 
next generation sequencing and gene expression measurement technology and its 
wide applications in research.

The symposium will focus on 70 lectures and a huge poster exhibition will be 
presented by internationally recognised experts in their field. The emphasis 
will be on unbiased, didactic information exchange. Internationally renown 
speakers will be participating in a lively and exciting programme enabling the 
valuable exchange of information in the qPCR and Next Generation Sequencing 
field. One third of the talks will be presented by selected invited speakers, 
one third will be selected from the submitted abstracts and one third will be 
presented by qPCR & NGS related company R&D representatives. All scientific 
contributions will be published in the Symposium Proceedings (ISBN to be 
announced).

Full papers from selected invited academic and industrial speakers and 
application notes from industrial speakers will be published in a METHODS 
special issue “Transcriptional Biomarkers”  edited by Michael W. Pfaffl 
(published January 2013).  At the meeting all participants will get  a free 
hard cover of this special Methods issue. Please have a look to our previous 
issue  => “The ongoing evolution of qPCR” METHODS special qPCR Vol 50 issue 4  
(April 2010)

Please register using the Internet based ConfTool registration and submission 
platform => http://registration.qPCR-NGS-2013.net

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Symposium Talk and Poster sessions:
http://sessions.qpcr-ngs-2013.net/

- Main Topic:   Molecular diagnostics
- Main Topic:   Next Generation Sequencing  (NGS)
- Main Topic:   Transcriptional Biomarkers
- High throughput analysis in qPCR
- Systems biology
- Single-cells diagnostics
- MIQE & QM strategies in qPCR
- non-coding RNAs - microRNA, siRNA, long non-coding RNAs
- Digital PCR  &  Nano-fluidics
- Pre-analytical Steps
- BioStatistics & BioInformatics
- qPCR & NGS data analysis
- Lunch Seminars:
   - qBASEplus - data analysis lunch seminar
   - GenEx - data analysis lunch seminar
   - NGS data analysis lunch seminars
   - more to be announced........


View our qPCR 2011 event trailer on YouTube => 
http://www.youtube.com/watch?v=cp8WwPyLW8Y

Download the latest Event Flyer => http://tinyurl.com/qPCR-NGS-2013

Please register using the Internet based ConfTool registration and submission 
platform => http://registration.qPCR-NGS-2013.net

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GenEx 5 -  A Powerful Tool For qPCR Data Analysis
Download a free trail version here => http://GenEx.gene-quantification.info

GenEx is a 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. View our webpage => 
http://GenEx.gene-quantification.info

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, Principal Component Analysis, 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.

Download a free trail version here => http://GenEx.gene-quantification.info

GenEx PDF user guides:
* GenEx user guide 
* GenEx user guide - Exiqon Wizard 
* GenEx user guide - Roche Wizard


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  to further scientists and friends who are interested in qPCR !

Best regards,
Michael W. Pfaffl
responsible Editor of the Gene Quantification Pages


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