qPCR / real-time PCR

qPCR newsletter - January 2012

2012-01-27T18:06:00.005+01:00

The January 2012 qPCR NEWS 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:

digital-PCR application papers - digital-PCR.gene-quantification.info
real-time PCR optimisation - optimisation.gene-quantification.info
GenEx version 5 - download a free trial version - GenEx.gene-quantification.info
follow our newsletter qPCRnews.gene-quantification.info

A MIQE Case Study — Effect of RNA Sample Quality and Reference Gene Stability on Gene Expression Data

2012-01-07T15:46:00.008+01:00

A MIQE Case Study — Effect of RNA Sample Quality and Reference Gene Stability on Gene Expression Data

by Sean Taylor, Bio-Rad Laboratories Canada, 1329 Meyerside Dr Mississauga, ON, L5T 1C9, and Marguerite Buchanan and Mark Basik, McGill University, Jewish General Hospital, Montreal, QC, published: December 15, 2011

Abstract - Real-time quantitative PCR (qPCR) has become the gold standard for validating DNA microarray data and is routinely used to determine gene expression differences between a wide variety of samples. The exquisite sensitivity of the technology permits the detection of a single copy of a target gene in a sample which has led to qPCR now being used in the clinical setting to diagnose infection and disease states. In an effort to standardize the design of the associated experiments, the minimum information for publication of quantitative real-time PCR experiments (MIQE) guidelines were published in 2009. In this study, we show how qPCR can lead to erroneous conclusions regarding differences in MCM7 gene expression between normal and tumor human breast cancer samples if the key steps set out in the MIQE guidelines are not followed.

Practical Course on Single-Cell Gene Expression Analysis

2011-12-22T07:59:00.002+01:00

EMBO Practical Course

Single-Cell Gene Expression Analysis

EMBL Heidelberg, GermanyMonday 19 March - Friday 23 March 2012

http://www.embl.de/training/events/2012/SIC12-01/index.html

EMBL Master Course

2011-12-22T07:57:00.002+01:00

EMBL Master Course:

Advanced qPCR Techniques for Publication Success: Following MIQE Recommendation

EMBL Heidelberg, Germany; Monday 2 July - Friday 6 July 2012 http://www.embl.de/training/events/2012/MIQ12-01/index.html

qPCR newsletter December 2011

2011-12-20T16:48:00.003+01:00

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:

qPCR efficiency calculation - sub-domain updated - http://Efficiency.gene-quantification.info
real-time PCR Cycler - sub-domain updated - http://CYCLERS.gene-quantification.info
GenEx version 5 - download a free trial version - http://GenEx.gene-quantification.info
MIQE qPCR APP for iOS and Android is available - http://MIQE.gene-quantification.info

GenEx version 5 for FREE download

2011-12-09T10:28:00.003+01:00

New GenEx 5 version offers advanced methods to analyze real-time qPCR data with simple clicks of the mouse.
GenEx is the leading software for processing and analysis of qPCR data. It provides a multitude of functionalities for the qPCR community, ranging from basic data editing and management to cutting-edge data analysis.
Near universal instrument compatibility - GenEx is already compatible with instruments from most of the main qPCR instrument producers and the list of compatible instruments keeps growing with the help and input from existing customers.
User-friendly interface - GenEx has a user friendly interface designed to be both easy to learn for a novice and flexible enough for the demands of a professional user.
Comprehensive pre-processing and data editing - Possibly the most important part of qPCR experiments is to pre-process the raw data for subsequent statistical analyses. The pre-processing steps need to be performed consistently in correct order and with confidence. GenEx contains comprehensive tools for data pre-processing and data editing.
Cutting-edge data analysis - GenEx offers cutting-edge data analysis tools, with GenEx Pro and Enterprise being the most complete versions. Current features include parametric and non-parametric statistical tests, clustering methods, principal component analysis, artificial neural networks, and much more. New features are continuously being added to GenEx with close attention to customer needs. In the GenEx forum you will find in depth information about experimental design, data analysis and GenEx features.

Tutorials:

qPCR NEWS - November 2011 - focus on mirtrons

2011-11-24T14:30:00.009+01:00

Our Nov 2011 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 main focus of this newsletter issue is:

* Introducing the mirtron - http://mirtron.gene-quantification.info

* MIQE qPCR APP for IOS and Android is available - http://MIQE.gene-quantification.info

If this newsletter is not displayed correctly by your email client, please use following http://qPCRnews.gene-quantification.info

BioTechniques - Future Methods in PCR

2011-11-19T17:46:00.004+01:00

BioTechniques - Future Methods in PCR

Leroy Hood, Carl Wittwer, Roopom Banerjee, Anna Berdine, and Junko Stevens
discuss the current limitations and opportunities in the development of PCR techniques.
In our 2011 Future Methods article series, BioTechniques presents in-depth interviews with the thought-leaders and methods developers in the fields of cell biology, genomics, proteomics, epigenetics, and microfluidics.
These articles will present the perspectives and thoughts of leading researchers, focusing on the technologies that are being developed to advance research efforts. These articles will provide new information and insights, and a glimpse into how methods and techniques might change in the years to come.

new MIQE APP available on Android Market

2011-11-07T20:15:00.003+01:00

ANDROID MARKET MIQE qPCR - Get help from a special team of experts in qPCR while on the move. MIQE - qPCR helps you in reviewing scientific works and checking your own experiments, when qPCR is involved. Check your project's compliance to MIQE in minutes, have all required references to hand, and follow qPCR events and news.
https://market.android.com/details?id=com.biorad.miqeqPCR

Gene-Expression Analysis Exploits More Technologies

2011-11-05T13:49:00.004+01:00

Gene-Expression Analysis Exploits More Technologies
To quantify the expression of specific genes, researchers can use a variety of techniques, including arrays, PCR, and high throughput sequencing. However, getting accurate results still depends on precisely carrying out these methods, even with increasingly user-friendly technologies. In fact, as more scientists study gene expression, the standards for analysis are growing more rigorous to ensure that only accurate data are published. Likewise, software has been keeping pace, helping researchers follow protocols and analyze their results.

By Mike May

2011-11-02T21:18:00.004+01:00

In search of better real-timePCR data

http://www.dddmag.com/article-In-Search-of-Better-Real-Time-PCR-10111.aspx

By Richard Kurtz, In Drug Discovery & Development - October 01, 2011

Real-time quantitative PCR (qPCR) has become the industry standard for the detection and quantification of nucleic acids. However, the lack of consensus among researchers on how to best perform and interpret qPCR experiments is a major hurdle for advancing the technology. This problem is exacerbated when insufficient experimental detail is given in published work, impeding the ability of others to accurately evaluate or replicate reported results.

qPCR NEWS - October 2011 - focus on MIQE and RefGenes

2011-10-13T10:22:00.005+02:00

qPCR NEWS - October 2011 - focus on MIQE and RefGenes

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 the October 2011 newsletter issue is:

MIQE Guidelines are entering "high impact" journals! - http://MIQE-press.gene-quantification.info
RefGenes - a unique tool to find suitable reference genes - http://www.refgenes.org - http://normalisation.gene-quantification.info
MIQE qPCR APP for iPhone, iPad and iPod - new updated version available - http://MIQE.gene-quantification.info
Join the qPCR 2011 US - http://www.qPCRsymposium.com

Follow the complete newsletter on http://qPCRnews.gene-quantification.info

LIFE SCIENCE TECHNOLOGIES - qPCR Innovations and Blueprints

2011-10-07T18:28:00.004+02:00

LIFE SCIENCE TECHNOLOGIES - qPCR Innovations and Blueprints PDF
By Chris Tachibana
http://www.sciencemag.org/

Quantitative PCR users can rapidly generate large amounts of high-quality data with new instruments and products made possible by microfluidics and miniaturization technology. These platforms are the tools for developing techniques that require extremely high throughput and sensitivity such as digital PCR and single-cell analysis. Researchers are adopting these methods to ask sophisticated questions about genetics and cancer biology as well as to develop novel research and diagnostic assays. As qPCR innovators explore new frontiers and everyday users venture into more complicated workflows, international groups of industry and academic partners are keeping us on the path of best practices. Two consortia (MIQE & SPIDIA) are generating guidelines on the qPCR process - from experimental design and pre-analysis sample collection, to processing data and publishing results. The guidelines are blueprints that ensure reproducibility, validity, and transparency.

Next meeting of the New England RNA Data (NERD) club

2011-10-06T17:32:00.005+02:00

The next meeting of the New England RNA Data (NERD) club will take place in Cannon Room at Harvard Medical School on Wednesday, October 26, 2011.
http://www.newenglandrna.org/resources/NERD_October_2011.pdf

Our speakers will be:

Sebastian Marquardt, Steve Buratowski Lab, Harvard Medical School
"Differential RNA Processing of Long Noncoding Transcript Species in Budding Yeast"
Michael W. Pfaffl, Physiology Weihenstephan, Technische Universität München, Germany
"MIQE Challenges and Solutions - Reliable microRNA Expression Profiling and Biomarker Development"
Rachel Fearns, Microbiology Department, Boston University School of Medicine
"Initiating Replication on A Linear RNA Genome: Non-templated Initiation by the Respiratory Syncytial Virus Polymerase"

We will have beverages starting at 5:30PM and talks from 6:00-7:30PM in Cannon Room, 1st floor in Building C of HMS quadrangle.
Following the talks, we will convene outside the Cannon Room for a reception from 7:30-8:30pm.
This meeting is sponsored by *Qiagen*
For more detailed information, visit our website http://www.newenglandrna.org

qPCR NEWS - September 2011 issue

2011-09-21T13:56:00.003+02:00

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

Update of real-time qPCR page - outstanding reviews and innovative qPCR papers - qPCR.gene-quantification.info
Recorded qPCR 2011 Event - 65 interesting talks from the qPCR 2011 event in Freising-Weihenstephan - eConference.qPCR2011.net
MIQE guidelines - online translation service to CHINESE, JAPANESE, KOREAN and RUSSIAN
MIQE qPCR APP for iPhone, iPad and iPod - new version 1.1 available - MIQE.gene-quantification.info
US qPCR Symposium in Santa Clara, CA - 1st to 4th November 2011 - EVENTS.gene-quantification.info

MIQE guidelines Online Translation Service

2011-09-18T10:08:00.005+02:00

MIQE guidelines Online Translation Service
Since September 2011 we provide a direct translation of the MIQE guidelines in

CHINESE => http://miqe-chinese.gene-quantification.asia
JAPANESE => http://miqe-japanese.gene-quantification.asia
KOREAN => http://miqe-korean.gene-quantification.asia
RUSSIAN => http://miqe-russian.gene-quantification.asia

Please try the link multiple times. It is an automatic translation service. It takes a bit!

Recognize this is an automatic and robotic based translation service, and therefore we provide NO guarantee about the automatic generated content. It should help the world wide qPCR community to understand the core content of the Gene Quantification pages and MIQE guidelines, but still the original language is the English version.

eConference - qPCR 2011 EVENT is now online available!

2011-09-15T08:13:00.004+02:00

Follow our qPCR 2011 eConference
http://eConference.qPCR2011.net
The qPCR 2011 eConference package contains 65 talks in high resolution and very good sound quality, including all presented slides from the qPCR Symposium 2011 in Freising-Weihenstephan.

The talks are grouped in the following scientific sessions: single-cell qPCR, Next Generation Sequencing, High throughput qPCR, MIQE guidelines, RNAi-miRNA, qPCR Data analysis, HRM, Circulating Nucleic Acids, Pre-analytical Steps, New qPCR Applications in Molecular Diagnostics, and more ... ...

You can follow the outstanding talks from international well recognized speakers in the field of gene quantification and expression profiling: S. Bustin, J. Vandesompele, M. Kubista, K. Livak, MW. Pfaffl, Dennis Lo, A. Stahlberg, J. Helemanns, B. Brenig, R. Loewe, K. Hasenstein, A. Raj, F. McCaughan, K. Knudtson, P. Zimmermann, A. Nitsche, P. Day, A. Tichopad, and a lot of company representatives.

With our new eConference streaming server a world-wide know-how transfer becomes possible in a fast and simple way! You can watch an in-depth demo presentation of our talk’s right here => http://eConference.qPCR2011.net

qPCR and MIQE Seminar Series

2011-09-15T07:56:00.006+02:00

As part of our customer education program, we have provided two recorded seminar series covering the topics of qPCR and MIQE. The recorded sessions are intended to provide a high level overview of these subject matters. We have kept the lessons concise so that you can enjoy a self-paced learning program.

http://www.sigmaaldrich.com/life-science/learning-center/customer-education/qpcr-miqe-seminar-series.html

qPCR NEWSLETTER - August 2011 - focus on digital-PCR

2011-08-11T10:32:00.009+02:00

Dear researcher,

Update - digital PCR - digital-PCR.gene-quantification.info
Update of webinars & multi-media presentations & animations - WEBINAR.gene-quantification.info
Update of qPCR talks from 2004 - 2011 - TALKS.gene-quantification.info
MIQE qPCR APP for iPhone, iPad and iPod - new version 1.1 available since 3rd August 2011 - MIQE.gene-quantification.info
US qPCR Symposium in Santa Clara, CA - 1st-4th November 2011 - EVENTS.gene-quantification.info

NEW MIQE-qPCR APP is online since 3rd August

2011-08-08T07:53:00.002+02:00

The new MIQE_qPCR APP update version 1.1 is online since 3rd August => http://itunes.apple.com/app/miqe-qpcr/id423650002?mt=8

What's New in Version 1.1

Now MIQE qPCR app is even more interactive. For the first time ever, checklists are optimized in real time: you can reach 100% for every project if you are MIQE compliant.
Checklists are specific for each project type: just click on the kind of nucleic acid you are working with, the checklists adapts instantly by removing unnecessary items (Reverse transcription items are not relevant if working only on DNA for instance).
Moreover, some items may not apply to your specific experiments. You can now remove them and have the most accurate MIQE compliancy.
References have been updated, so you can keep in touch with latest MIQE related literature, symposium updates and more.
Last but not least: Export is available. You can archive the state of your project whenever you want, share it with your colleagues, and store it.
Suggestions collected from experts at the qPCR 2011 symposium in Freising were included.

Go, Go Gadgets - Research gets APP happy

2011-07-25T11:36:00.001+02:00

Go, Go Gadgets
July 13 2011 by Katia Caporiccio
For researchers working in life science, the MIQE qPCR app for iPhone and iPad, sponsored by Bio-Rad, provides resources and checklists needed to ensure MIQE (Minimum Information for Publication of Quantitative Real-Time qPCR Experiments) compliance for qPCR experiments. “In this day and age, everybody has his phone with him at all times,” said Rachel Scott, senior product manager for Gene Expression at Bio-Rad (Hercules, CA). “The immediacy helps with capturing the information as it occurs, on the fly.”

Research gets APP happy
06/23/2011 by Lisa Grauer
These two new iPhone and iPad apps promise to advance your research more than any summer intern.
Bio-Rad’s new MIQE qPCR app also comes equipped with lab tools including extensive qPCR reference information and checklists that allow researchers to ensure MIQE compliance for their qPCR experiments.
“qPCR is a common lab technique used by most life science researchers today, but not everyone conducting PCR is using the technique in a way that ensures its correct interpretation,” said Rachel Scott, Bio-Rad senior product manager. “As a result of misinterpretation of qPCR data, significant scientific conclusions have been retracted for inaccuracies.”
Developed by real-time PCR (qPCR) experts Michael W. Pfaffl, professor of molecular physiology at Technische Universität München, and Afif Abdel Nour, associate professor of nurigenomics at the Institut Polytechnique LaSalle Beauvais, the MIQE qPCR app helps researchers achieve accuracy, transparency, and reproducibility in their qPCR experiments by allowing them to monitor MIQE compliance via color-coded checklists and progress bars. The app also provides expert advice through direct links to MIQE-related publications and email addresses of qPCR experts.

our latest paper - Quantification noise in single cell experiments

2011-07-13T14:57:00.002+02:00

Quantification noise in single cell experiments

M. Reiter1, B. Kirchner2, H. Müller3, C. Holzhauer3, W. Mann3 and M. W. Pfaffl2

1) BioEPS GmbH, Lise-Meitner-Strasse 30, 85354 Freising

2) Physiology Weihenstephan, TUM – Technische Universität München, Weihenstephaner Berg 3, 85354 Freising

3) Beckman Coulter Biomedical GmbH, Sauerbruchstraße 50, 81377 Munich, Germany

Abstract: In quantitative single-cell studies, the critical part is the low amount of nucleic acids present and the resulting experimental variations. In addition biological data obtained from heterogeneous tissue are not reflecting the expression behaviour of every single-cell. These variations can be derived from natural biological variance or can be introduced externally. Both have negative effects on the quantification result. The aim of this study is to make quantitative single-cell studies more transparent and reliable in order to fulfil the MIQE guidelines at the single-cell level. The technical variability introduced by RT, pre-amplification, evaporation, biological material and qPCR itself was evaluated by using RNA or DNA standards. Secondly, the biological expression variances of GAPDH, TNFα, IL-1β, TLR4 were measured by mRNA profiling experiment in single lymphocytes. The used quantification setup was sensitive enough to detect single standard copies and transcripts out of one solitary cell. Most variability was introduced by RT, followed by evaporation, and pre-amplification. The qPCR analysis and the biological matrix introduced only minor variability. Both conducted studies impressively demonstrate the heterogeneity of expression patterns in individual cells and showed clearly today's limitation in quantitative single-cell expression analysis.
http://nar.oxfordjournals.org/content/early/2011/07/08/nar.gkr505.abstract

more than 1,000 downloads of MIQE_qPCR APP

2011-06-28T16:11:00.003+02:00

We are pleased to tell you that we have reached more then 1,000 downloads of the MIQE_qPCR APP from 39 countries:

USA 367 downloads

France 78 downloads

UK 54 downloads

China 47 downloads

Germany 38 downloads

Australia 37 downloads

Canada 36 downloads

Mexico 35 downloads

Italy 31 downloads

Korea 31 downloads

other 29 countries 273 downloads

http://www.facebook.com/MIQE.qPCR

Follow our qPCR 2011 eConference

2011-05-31T15:07:00.007+02:00

Follow our qPCR 2011 eConference
http://econference.qpcr2011.net
The qPCR 2011 package contains 65 talks from the qPCR Symposium 2011 in Freising-Weihenstephan

Topics: Single Cell qPCR, Next Generation Sequencing, High throughput qPCR, MIQE guidelines, RNAi-miRNA, qPCR Data analysis, HRM, Circulating Nucleic Acids, Pre-analytical Steps, New qPCR Applications in Molecular Diagnostics, and more ... ...

Bio-Rad Introduces the First MIQE qPCR App, Now Available at the Apple App Store

2011-05-25T07:54:00.005+02:00

Hercules, CA — May 23, 2011 — The first MIQE qPCR app, sponsored by Bio-Rad Laboratories, Inc., is now available for free through Apple's App Store. The new app provides researchers with resources and checklists needed to ensure MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) compliance for their qPCR experiments.

"qPCR is a common lab technique used by most life science researchers today, but not everyone conducting PCR is using the technique in a way that ensures its correct interpretation," said Rachel Scott, PhD, Bio-Rad Senior Product Manager of the Gene Expression Division. "As a result of misinterpretation of qPCR data, significant scientific conclusions have been retracted for inaccuracies. The MIQE qPCR app gives users the ability to check off applicable MIQE attributes, which in turn increases the users' and the scientific community's confidence in the project's validity."

Developed by real-time PCR (qPCR) experts Michael W. Pfaffl, a professor at Technische Universität München, and Afif Abdel Nour, an associate professor at the Institut Polytechnique LaSalle Beauvais, the MIQE qPCR app helps researchers achieve accuracy, transparency, and reproducibility in their qPCR experiments by allowing them to:

Monitor MIQE compliance — Color-coded checklists distinguish between essential and desirable MIQE requirements; a visual indication of progress is provided

Get expert advice — Users can link directly to MIQE-related publications or direct their qPCR or MIQE-related questions to the experts listed via email

Stay up to date — The most current qPCR resources, references, and news are readily available

Impact the future of MIQE guidelines — Users can send their completed project checklists to the worldwide MIQE guideline database where the information will be used to further refine the MIQE guidelines

Bio-Rad sponsored the MIQE qPCR app as part of its commitment of giving scientists tools to optimize and validate their qPCR experiments according to MIQE guidelines.

The MIQE qPCR App runs on Apple's iPhone, iPod Touch, or iPad and can be downloaded here.

About Bio-Rad
Bio-Rad Laboratories, Inc. (NYSE: BIO and BIOb) has remained at the center of scientific discovery for more than 50 years, manufacturing and distributing a broad range of products for the life science research and clinical diagnostic markets. The company is renowned worldwide among hospitals, universities, and major research institutions, as well as biotechnology and pharmaceutical companies for its commitment to quality and customer service. Founded in 1952, Bio-Rad is headquartered in Hercules, California, and serves more than 100,000 research and industry customers worldwide through its global network of operations. The company employs more than 6,800 people globally and had revenues exceeding $1.9 billion in 2010. For more information, please visit http://www.biorad.com

For more information contact:
Rachel Scott
Bio-Rad Laboratories, Inc.
510-741-6336
Rachel_Scott@bio-rad.com

Ken Li
Chempetitive Group
312-997-2436
kli@chempetitive.com

Video Tutorial: MIQE and your qPCR data

2011-05-20T10:15:00.002+02:00

Video Tutorial: MIQE and your qPCR data

May 19, 2011 by Bio-Rad

In this video, Dr. Sean Taylor, Field Applications Specialist, Bio-Rad Laboratories, demonstrates how sample quality and reference gene selection effect data analysis and interpretation in real-time quantitative PCR (qPCR) experiments. The presentation is in accordance with the previously published MIQE guidelines.For enhanced viewing, click on the full-screen mode button on the bottom right hand corner of the video.

Evaluierung der qPCR

2011-05-04T10:07:00.002+02:00

Die Real-Time-RT-PCR-Datenanalyse im Fokus der MIQE-Richtlinie
BIOspektrum, Mai 2011, 295-297
http://www.biospektrum.de/artikel/1066992
MICHAEL W. PFAFFL & IRMGARD RIEDMAIER
LEHRSTUHL FÜR PHYSIOLOGIE, WISSENSCHAFTSZENTRUM WEIHENSTEPHAN FÜR ERNÄHRUNG, LANDNUTZUNG & UMWELT, TU MÜNCHEN
Die MIQE-Richtlinie wurde 2009 von einer Gruppe internationaler Wissenschaftler ins Leben gerufen, um die Qualität, die Richtigkeit sowie die Zuverlässigkeit der gewonnenen qPCR-Ergebnisse im Labor und in der
wissenschaftlichen Literatur zu steigern.
The MIQE guidelines were established 2009 by a group of international scientist to improve the quality, the accuracy, and the reliability of the generated quantitative PCR results in the lab and in the scientific literature.

Follow our new facebook MIQE.qPCR community

2011-04-12T16:43:00.012+02:00

MIQE_qPCR APP for iPhone, iPad and iPod

2011-04-08T08:06:00.003+02:00

MIQE_qPCR - download our new MIQE qPCR APP for iPhone, iPad and iPod - developed by LaSalle Beauvais University, Technical University of Munich and Bio-Rad - http://goo.gl/fb/E4pyx

PCR NEWS - February 2011

2011-02-23T13:58:00.001+01:00

updated summary of real-time PCR cyclers => http://cyclers.gene-quantification.info
update of interesting webinars => http://webinar.gene-quantification.info
qPCR 2011 Event => Final Announcement
qPCR 2011 Symposium Agenda presenting 71 talks and 90 posters

qPCR 2011 Event Agenda is online

2011-02-16T08:01:00.009+01:00

Over 170 contributions were submitted and 71 talks and 90 posters are approved to be presented at the symposium. Symposium agenda is online http://agenda.qpcr2011.net/ and http://posters.qpcr2011.net/

qPCR 2011 Agenda PDF with timing in available => http://agendapdf.qpcr2011.net/
Join our next qPCR Event - http://registration.qpcr2011.net/

qPCR Event exhibition
28th -30th March is FULLY booked by 35 biotec companies - http://exhibition.qpcr2011.net/

qPCR Event Sponsors:
LEAD SPONSORS: Roche & Agilent Technologies
GOLD SPONSORS: Bio-Rad, Biosearch Technologies, Qiagen, IDT, Life Technologies, Sigma LifeScience, ThermoFisher Scientific, Exiqon, Nanostring, Fluidigm
SILVER SPONSORS: Premier Biosoft International, Wafergen, Eurogentec, TwistDx
http://sponsors.qpcr2011.net/

qPCR NEWS - Jan 2011 - Data Analysis & Bioinformatics & qPCR efficiency

2011-01-29T10:09:00.007+01:00

Dear researcher,
dear Gene Quantification page reader,

Our newsletter informs about the latest news in quantitative real-time PCR (qPCR and qRT-PCR), which are compiled and summarised on the Gene Quantification homepage. The focus of this newsletter issue is:
- update of new publications in qPCR Biostatistics & Bioinformatics - http://bioinformatics.gene-quantification.info/
- update of new publications in qPCR efficiency calculation - http://efficiency.gene-quantification.info/
- qPCR 2011 Event - Final Call for TALK and POSTER abstracts - http://call.qpcr2011.net/
=> abstract submission deadline elongated till 10th Feb. 2011
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Data Analysis and BioInformatics in real-time qPCR
How to do successful gene expression analysis using real-time PCR
Stefaan Derveaux, Jo Vandesompele, Jan Hellemans; Methods Vol 50, Issue 4, April 2010, in The ongoing Evolution of qPCR edited by Michael W. Pfaffl, Pages 227-230

Reverse transcription quantitative PCR (RT-qPCR) is considered today as the gold standard for accurate, sensitive and fast measurement of gene expression. Unfortunately, what many users fail to appreciate is that numerous critical issues in the workflow need to be addressed before biologically meaningful and trustworthy conclusions can be drawn. Here, we review the entire workflow from the planning and preparation phase, over the actual real-time PCR cycling experiments to data-analysis and reporting steps. This process can be captured with the appropriate acronym PCR: plan/prepare, cycle and report. The key message is that quality assurance and quality control are essential throughout the entire RT-qPCR workflow; from living cells, over extraction of nucleic acids, storage, various enzymatic steps such as DNase treatment, reverse transcription and PCR amplification, to data-analysis and finally reporting.
Further papers added => http://www.gene-quantification.de/bioinf-subpage3.html

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Determination of real-time PCR amplification efficiency
Individual samples generate different and individual fluorescence histories in kinetic RT-PCR. The shapes of amplification curves differ in the steepness of any fluorescence increase and in the absolute fluorescence levels at plateau depending on background fluorescence levels. The PCR efficiency has a major impact on the fluorescence history and the accuracy of the calculated expression result and is critically influenced by PCR reaction components
More info here => http://efficiency.gene-quantification.info/

Experimental comparison of relative RT-qPCR quantification approaches for gene expression studies in poplar.
Regier N & Frey B.; BMC Mol Biol. 2010 11: 57, 8 pages

BACKGROUND: RT-qPCR is a powerful tool for analysing gene expression. It depends on measuring the increase in fluorescence emitted by a DNA-specific dye during the PCR reaction. For relative quantification, where the expression of a target gene is measured in relation to one or multiple reference genes, various mathematical approaches are published. The results of relative quantification can be considerably influenced by the chosen method.
RESULTS: We quantified gene expression of superoxide dismutase (SOD) and ascorbate peroxidase (APX) in the roots of two black poplar clones, 58-861 and Poli, which were subjected to drought stress. After proving the chosen reference genes actin (ACT), elongation factor 1 (EF1) and ubiquitin (UBQ) to be constantly expressed in the different watering regimes, we applied different approaches for relative quantification to the same raw fluorescence data. The results obtained using the comparative Cq method, LinRegPCR, qBase software and the Pfaffl model showed a good correlation, whereas calculation according to the Liu and Saint method produced highly variable results. However, it has been shown that the most reliable approach for calculation of the amplification efficiency is using the mean increase in fluorescence during PCR in each individual reaction. Accordingly, we could improve the quality of our results by applying the mean amplification efficiencies for each amplicon to the Liu and Saint method.
CONCLUSIONS: As we could show that gene expression results can vary depending on the approach used for quantification, we recommend to carefully evaluate different quantification approaches before using them in studies analysing gene expression.
New added publications => http://efficiency.gene-quantification.info/
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qPCR 2011 Event - 5th international qPCR Symposium & Industrial Exhibition & Application Workshop
28th March - 1st April 2011
in Freising-Weihenstephan, Technical University of Munich, Weihenstephan, Germany

qPCR 2011 Event - Final Call for TALK and POSTER abstracts - http://call.qpcr2011.net/
Please submit your abstract here => http://submission.qpcr2011.net/

The symposium will focus on 60-70 lectures and more than 100 posters will be presented by internationally recognised experts in their field. The emphasis will be on unbiased, didactic information exchange. Internationally reknown speakers will be participating in a lively and exciting programme enabling the valuable exchange of information in the qPCR 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 related company R&D representatives. All scientific contributions will be published in the qPCR 2011 Symposium Proceedings.
Please register here => http://registration.qpcr2011.net/
Download 1st qPCR 2011 symposium announcement => http://tinyurl.com/CALLqPCR2011

VERY VERY COOL - Zheng Lab - Bad Project (Lady Gaga parody)

2011-01-28T10:28:00.002+01:00

http://www.youtube.com/watch?v=Fl4L4M8m4d0

FINAL CALL for TALK & POSTER submission qPCR 2011 Event

2011-01-28T08:06:00.007+01:00

FINAL CALL for TALK & POSTER submission qPCR 2011 Event
Submission deadline and early Registration period will end on 31st January 2011 http://call.qpcr2011.net/
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qPCR 2011 Event - 5th international qPCR Symposium & Industrial Exhibition & Application Workshop
28th March - 1st April 2011, in Freising-Weihenstephan, Technical University of Munich, Physiology-Weihenstephan, Germany
Download CALL => http://tinyurl.com/CALLqPCR2011

Sessions:

Molecular diagnostics in single-cells
High throughput analysis in qPCR & Next Generation Sequencing
MIQE and QM strategies in qPCR
small RNAs qPCR - microRNA and siRNA Applications
Digital PCR & Nano-fluidics
Pre-analytical Steps
qPCR BioStatistics & BioInformatics

Please submit your contribution here => http://submission.qpcr2011.net/
Please register here => http://registration.qpcr2011.net/
=> Abstract submission deadline shifted to 10th February 2011

looking forward to meet you at the symposium in March 2011

Michael Pfaffl

qPCR NEWS Dec 2010 - focus on RNA and DNA quality control for qPCR

2010-12-22T12:43:00.006+01:00

qPCR NEWS Dec 2010 - focus on RNA and DNA quality control for qPCR

Dear researcher,

updates on RNA and DNA quality control http://rna-integrity.gene-quantification.info/
updates on MIQE media review - http://miqe-press.gene-quantification.info/
qPCR 2011 Event - Call for TALK and POSTER abstracts - http://call.qpcr2011.net/
qPCR Application Workshops Jan/Feb 2011 - http://site.bioeps.com/index.php?option=com_seminar&Itemid=6
Full NEWSLETTER - http://qPCRnews.gene-quantification.info/

An Interview with MIQE Guru and qPCR Expert Stephen Bustin

2010-12-13T17:31:00.000+01:00

An Interview with MIQE Guru and qPCR Expert Stephen Bustin

qPCR 2011 Event - Industrial Exhibition almost fully booked

2010-12-07T07:47:00.003+01:00

http://www.qPCR2011.net/
The focus of the qPCR 2011 Event will be on:
Molecular Diagnostics: from single-cells to Next Generation Sequencing 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 Talk & Poster Sessions, Industrial Exhibition and associated three qPCR Application Workshops offer an overview of the present knowledge and future developments in qPCR and gene expression measurement technology and its wide applications.The symposium will focus on 60-70 lectures and more than 100 posters will be presented by internationally recognised experts in their field. The emphasis will be on unbiased, didactic information exchange. Internationally reknown speakers will be participating in a lively and exciting programme enabling the valuable exchange of information in the qPCR field.

Main topic: Molecular diagnostics in single-cells
Main topic: High throughput analysis in qPCR & NGS (Next Generation Sequencing)
MIQE and QM strategies in qPCR
small RNAs qPCR - microRNA and siRNA
Digital PCR & Nano-Fluidics
Pre-analytical steps
BioStatistics & BioInformatics

If your company is interested to participate at the qPCR 2011 Event => BOOK NOW !
The Industrial Exhibition is almost fully booked => http://exhibition.qpcr2011.net/

Professional event organization: Dr. Martina Reiter, BioEPS GmbH, Freising, Germany Martina.Reiter@BioEPS.com www.bioEPS.com

Next Basic qPCR Application Workshop 17-19 January 2011

2010-11-14T10:18:00.005+01:00

Introductory course consisting of theoretical and practical parts. Issues: qPCR introduction, assay optimization, sampling and extraction methods, data analysis and quantification strategies. Language - English
Date: 17 - 19 January 2011
Location: BioEPS GmbH, Lise-Meitner-Strasse 30, 85354 Freising, Germany (very close to Munich airport MUC)
Link => http://site.bioeps.com/index.php?option=com_seminar&Itemid=6

Day 1:

Basic PCR theory and applications
Detection Chemistry (SYBR Green I, TaqMan, Molecular Beacons...etc)
Different instrument platforms and their typical uses
Primer Design, Probe design and optimization
Basic data handling and analysis

Day 2:

Experimental setup and how it influences qPCR data
Nucleic acid extraction
cDNA synthesis and pre-amplification
Introduction to specific qPCR applications: genotyping, SNP analysis, HRM

Day 3:

Introduction to quantification principles
Quantification strategies:
Absolute quantification
Relative quantification
qPCR efficiency
Strategies for normalization of qPCR data

About the tutors:
Dr. Michael Pfaffl is one of the leading experts in qPCR data analysis, specialiced on relative quantification strategies. He is author of many highly influental publications.
Dr. Martina Reiter is qPCR specialist with focus on experiment setup and many years of experience in the optimization of qPCR experiments.

Basic qPCR eSeminar

2010-11-13T17:02:00.000+01:00

qPCR Basics eSeminar Module is an introductory course for beginners in quantitative PCR analysis. Participants get a general overview about qPCR technology, primer design and assay optimization, so that they could start with first qPCR analysis.The course is done in cooperation with different companies to give the participants a general and independent survey about existing technologies and products.
Issues:
- Basic PCR theory and applications
- Detection chemistry (SYBR Green I, TaqMan, Molecular Beacons...etc)
- Different instrument platforms and their typical uses
- Primer design, probe design
- PCR assay optimization
Available on http://eseminars.bioeps.com/shop
Watch demo on http://eseminars.bioeps.com/player/?action=demo

The ongoing evolution of qPCR

2010-11-13T17:00:00.002+01:00

Follow 50 exciting talks from the qPCR 2010 Event in Vienna on - http://econference.bioeps.com/ or http://evolution.gene-quantification.info/

With our new eSeminars and eConference tool a world-wide know-how transfer becomes possible in a fast and simple way.http://eSeminars.bioeps.com/player/?action=demo

The ongoing evolution of qPCRThe polymerase chain reaction (PCR) is usually described as a simple, sensitive and rapid technique that uses oligonucleotide primers, dNTPs and a heat stable Taq polymerase to amplify DNA. It was invented by Kary B. Mullis and co-workers in the early eighties, who were awarded the 1993 Nobel Prize for chemistry for this discovery. With the discovery of real-time PCR in the nineties the method took an important hurdle towards becoming “fully quantitative”. The addition of an initial reverse-transcription (RT) step produced the complementary RT-PCR, a powerful means of amplifying any type of RNA. Today quantitative PCR (qPCR) is widely used in research and diagnostics, with numerous scientists contributing to the pre-eminence of PCR in a huge range of DNA-, RNA- (coding and non-coding) or protein- (immuno- or proximity ligation assay qPCR) based applications. Soon the PCR was regarded as the “gold standard” in the quantitative analysis of nucleic acid, because of its high sensitivity, good reproducibility, broad dynamic quantification range, easy use and reasonable good value for money.qPCR has substantial advantages in quantifying low target copy numbers from limited amounts of tissue or identifying minor changes in mRNA or microRNA expression levels in samples with low RNA concentrations or from single cells analysis. The extensive potential to quantify nucleic acids in any kind of biological matrix has kept qPCR at the forefront of extensive research efforts aimed at developing new or improved applications. But are qPCR and its associated quantification workflow really as simple as we assume?It is essential to have a comprehensive understanding of the underlying basic principles, error sources and general problems inherent with qPCR and RT-qPCR. This rapidly reveals the urgent need to promote efforts towards more reproducible, sensitive, truly quantitative and, ultimately, more biologically valid experimental approaches. Therefore, the challenge is to develop assays that meet current analytical requirements and anticipate new problems, for example in novel biological matrices or for higher throughput applications. Unfortunately, we are far from having developed optimal workflows, the highest sensitivity, the best RNA integrity metrics or the ultimate real-time cycler, all of which are indispensable for optimal PCR amplification and authentic results. The qPCR research community still aims to improve and evolve, which brings to the topic of this PCR special issue - The ongoing evolution of qPCR.

qPCR 2011 Event in Freising Weihenstephan

2010-10-27T17:45:00.009+02:00

http://www.qpcr2011.net/
The focus of the qPCR 2011 Event will be on:
Molecular Diagnostics: from single-cells to Next Generation Seqeuncing 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 Application Workshops offer an overview of the present knowledge and future developments in qPCR and gene expression measurement technology and its wide applications.The symposium will focus on 60-70 lectures and more than 100 posters will be presented by internationally recognised experts in their field. The emphasis will be on unbiased, didactic information exchange. Internationally reknown speakers will be participating in a lively and exciting programme enabling the valuable exchange of information in the qPCR field.

Main topic: Molecular diagnostics in single-cells
Main topic: High throughput analysis in qPCR & NGS (Next Generation Sequencing)
MIQE and QM strategies in qPCR
small RNAs qPCR - microRNA and siRNA
Digital PCR & Nano-Fluidics
Pre-analytical steps
BioStatistics & BioInformatics

The scientific organization is managed by international well-known scientists in the field of real-time PCR:

Stephen Bustin, Prof. of Molecular Science QM, School of Medicine, London, UK
Mikael Kubista, Prof. of Biotechnology, TATAA Biocenter, Sweden
Jo Vandesompele, Prof. at the Center of Medical Genetics, University of Ghent, Belgium
Heinrich H.D. Meyer, Prof. of Physiology, Weihenstephan, Germany
Michael W. Pfaffl, Reader in Physiology, Weihenstephan, Germany, scientific coordinator of the Symposium and the Application Workshops
Event organization: Dr. Martina Reiter BioEPS GmbH Freising Martina.Reiter@BioEPS.com

Single-Cell Challenges - GEN article

2010-10-19T11:54:00.003+02:00

Utilizing qPCR for single-cell analysis remains a significant hurdle, according to Martina Reiter, Ph.D., CEO of BioEPS. “Although single-cell analysis is not new, we still don’t have a final solution as to how to handle it. The biggest problem is technical variance during the experimental steps, especially in sampling analysis and subsequently data analysis.”
“Often samples are generated from laser capture microdissection and flow cytometry sorting. Sample amounts are very small and variances are huge. When you don’t pay attention in the beginning to such variances, downstream processes are even more inaccurate.”
What about solutions? “We are at the beginning of working on this and are continuing to determine the right way to do it,” Dr. Reiter said. “Substantial improvements in the analysis steps are needed. You cannot use standard analysis methods; they need to be modified for single-cell qPCR. But it is still difficult to modify so that technical variances are reduced as much as possible. Despite this, qPCR technology is sensitive enough to detect specific genes out of one cell.”
The MIQE guidelines are a newly proposed set of suggestions to standardize and document qPCR. “MIQE does not cover specific single-cell analysis so far, and qPCR advances need to be made in documenting and determining RNA integrity, in how to handle the necessity of preamplification, in the ability to more accurately sample cells, and in downstream analysis methodologies.”
Recognizing the importance of tackling these issues now will especially impact the future of therapeutics. “Ultimately, these problems will be solved,” Dr. Reiter predicted. “For the present, we need to realize that while qPCR for single-cell analysis is possible, especially technically, the larger issues of experimental design and variation remain and should be addressed.”
Link to article

Join our microRNA qPCR application workshop in November

2010-09-29T16:20:00.005+02:00

Theoretical and practical parts about small silencing RNA (miRNA, siRNA) with focus on quantitative microRNA analysis. For experienced users in miRNA and PCR.
Language: In English

Date: 8-9 November 2010
Location: BioEPS GmbH, Lise-Meitner-Strasse 30, 85354 Freising, Germany (very close to Munich airport MUC)
Link => http://site.bioeps.com/index.php?option=com_seminar&Itemid=6
Day 1: Introduction, microRNA and RNA Quality
- Introduction siRNA, microRNA, piRNA, RNAi: biogenesis, action and applications
- RNA Isolation: setting up a miRNA based experiment
- microRNA Quantity and Quality Control: methods and applications
- RT-qPCR: priciples of RT and qPCR, difference between enzymes and primers
- Optimization strategies for miRNA
Practical Part: real-time miRNA-qPCR Experiment
Day 2: Normalization and Quantification
- Data analysis: principles of qPCR softwares, algorithms and melt curve analysis
- Quantification strategies: absolute and relative quantification strategies
- Normalization with miRNA: how to choose a good reference
Practical Part: real-time miRNA-qPCR Experiment

Making the most of MIQE - MIQE precis

2010-09-23T08:56:00.010+02:00

Making the most of MIQE
The Editorial Board of BMC Molecular Biology endorse a new set of essential MIQE-light guidelines for the reporting of quantitative PCR data: "MIQE precis", and provide guidance for the suitability of pure reference gene papers to the journal.

MIQE precis: Practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments
Stephen A Bustin, Jean-Francois Beaulieu, Jim Huggett, Rolf Jaggi, Frederick SB Kibenge, Pal A Olsvik, Louis C Penning and Stefan Toegel

BMC Molecular Biology 2010; Published: 21 September 2010

The conclusions of thousands of peer-reviewed publications rely on data obtained using fluorescence-based quantitative real-time PCR technology. However, the inadequate reporting of experimental detail, combined with the frequent use of flawed protocols is leading to the publication of papers that may not be technically appropriate. We take the view that this problem requires the delineation of a more transparent and comprehensive reporting policy from scientific journals. This editorial aims to provide practical guidance for the incorporation of absolute minimum standards encompassing the key assay parameters for accurate design, documentation and reporting of qPCR experiments (MIQE precis) and guidance on the publication of pure 'reference gene' articles.

All news about the MIQE initiative => http://MIQE-press.gene-quantification.info

An Interview with MIQE Guru and qPCR Expert Stephen Bustin

2010-09-22T17:26:00.001+02:00

seen on - European Biotechnologist
Stephen Bustin obtained his Ph.D. from Trinity College, University of Dublin in molecular genetics in 1983. He is currently Professor of Molecular Science at Barts and the London School of Medicine and Dentistry where he aims to apply his research in a more translational setting by doing basic research and clinical practice in colorectal cancer. Bustin’s priority research aim is to translate his lab’s distinctive approach for predicting metastatic behaviour of colorectal cancer into a practical and robust assay for cancer prognosis that is not just more accurate than anatomically-based staging but can identify candidate therapeutic targets in metastatic colorectal cancers.
In a seminal paper published in Clinical Chemistry, Stephen Bustin et al. published a new set of guidelines that describe the minimum information necessary for evaluation of quantitative real-time polymerase chain reaction experiments. Following the Minimum Information for publication of Quantitative real-time PCR Experiments (MIQE) guidelines will encourage better experimental practice, allowing more reliable and unequivocal interpretation of quantitative PCR results.
=> follow the interview here

The Future of qPCR: Best practices, Standardization, and the MIQE Guidelines

2010-09-22T09:29:00.003+02:00

The Future of qPCR: Best practices, Standardization, and the MIQE Guidelines
Thursday, September 30, 2010 - 12 noon Eastern, 9 a.m. Pacific, 4 p.m. GMT
Quantitative polymerase chain reaction (qPCR) has emerged as a powerful tool in molecular biology laboratories, both in research and in diagnostic settings. Even as qPCR grows in popularity, it is being recognized that there are some challenges associated with the technology, particularly with respect to reproducibility within and between laboratories. Fortunately, many of these limitations can be addressed through a standardized set of best practices. Using the recently published MIQE guidelines as a foundation, our expert panel will address the best practices of qPCR, with the goal of providing researchers with more consistent and reliable data.

During the webinar, the panelists will:

provide an overview of the MIQE guidelines
address qPCR applications and primary challenges
outline best practices and assay design to get the best out of your qPCR
describe the essential quality control steps, including nucleic acid quantification
answer your questions during the live Q&A session

Participants:

Stephen A. Bustin, Ph.D.; Queen Mary, University of London; London, UK
Gregory L. Shipley, Ph.D.; University of Texas Health Science Center at Houston; Houston, TX
Manju R. Sethi; Thermo Fisher Scientific; Wilmington, DE

Review: Normalization strategies for microRNA profiling

2010-09-15T12:50:00.007+02:00

Normalization strategies for microRNA profiling experiments: a 'normal' way to a hidden layer of complexity?
Meyer SU, Pfaffl MW, Ulbrich SE
Biotechnol Lett. 2010 Aug 12
Physiology Weihenstephan, ZIEL Research Center for Nutrition and Food Sciences, Technische Universität München, Weihenstephaner Berg 3, 85354, Freising, Germany

Abstract
MicroRNA (miRNA) profiling is a first important step in elucidating miRNA functions. Real time quantitative PCR (RT-qPCR) and microarray hybridization approaches as well as ultra high throughput sequencing of miRNAs (small RNA-seq) are popular and widely used profiling methods. All of these profiling approaches face significant introduction of bias. Normalization, often an underestimated aspect of data processing, can minimize systematic technical or experimental variation and thus has significant impact on the detection of differentially expressed miRNAs. At present, there is no consensus normalization method for any of the three miRNA profiling approach. Several normalization techniques are currently in use, of which some are similar to mRNA profiling normalization methods, while others are specifically modified or developed for miRNA data. The characteristic nature of miRNA molecules, their composition and the resulting data distribution of profiling experiments challenges the selection of adequate normalization techniques. Based on miRNA profiling studies and comparative studies on normalization methods and their performances, this review provides a critical overview of commonly used and newly developed normalization methods for miRNA RT-qPCR, miRNA hybridization microarray, and small RNA-seq datasets. Emphasis is laid on the complexity, the importance and the potential for further optimization of normalization techniques for miRNA profiling datasets.

qPCR NEWS - August 2010 - focus on Copy Number Variations

2010-08-05T13:38:00.004+02:00

qPCR NEWS - August 2010 - focus on Copy Number Variations

UPDATE - Copy Number Variations => http://cnv.gene-quantification.info
eConference with high quality 50 talks presented at the "International qPCR 2010 Symposium in Vienna"
View our eSeminar and eConference demo videos http://eseminars.bioeps.com/player/?action=demo
qPCR Symposium USA in November 2010 => http://events.gene-quantification.info/
qPCR Application Workshops => http://workshops.gene-quantification.info

GenEx offers advanced methods to analyze real-time qPCR data

2010-07-28T15:03:00.007+02:00

GenEx offers advanced methods to analyze real-time qPCR data with simple clicks of the mouse.
GenEx is the leading software for processing and analysis of qPCR data. It provides a multitude of functionalities for the qPCR community, ranging from basic data editing and management to cutting-edge data analysis.
Near universal instrument compatibility - GenEx is already compatible with instruments from most of the main qPCR instrument producers and the list of compatible instruments keeps growing with the help and input from existing customers.
User-friendly interface - GenEx has a user friendly interface designed to be both easy to learn for a novice and flexible enough for the demands of a professional user.
Comprehensive pre-processing and data editing - Possibly the most important part of qPCR experiments is to pre-process the raw data for subsequent statistical analyses. The pre-processing steps need to be performed consistently in correct order and with confidence. GenEx contains comprehensive tools for data pre-processing and data editing.
Cutting-edge data analysis - GenEx offers cutting-edge data analysis tools, with GenEx Pro and Enterprise being the most complete versions. Current features include parametric and non-parametric statistical tests, clustering methods, principal component analysis, artificial neural networks, and much more. New features are continuously being added to GenEx with close attention to customer needs. In the GenEx forum you will find in depth information about experimental design, data analysis and GenEx features.
Tutorials:

Real Time QPCR Data Analysis Tutorial (part 2 relative quantification)

2010-07-27T15:45:00.001+02:00

Real Time QPCR Data Analysis Tutorial (part 1 absolute quantification)

2010-07-27T15:44:00.000+02:00

MIQE Guidelines on baidu.com

2010-07-26T08:56:00.000+02:00

MIQE Guidelines on baidu.com
MIQE press review => http://miqe-press.gene-quantification.info/

qPCR NEWS - July 2010 - focus on PCR efficiency

2010-07-22T16:16:00.002+02:00

qPCR NEWS - July 2010 - focus on PCR efficiency
http://qPCRnews.gene-quantification.info/

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

Update on qPCR efficiency estimation - Find lots of useful papers here => http://efficiency.gene-quantification.info
eConference with high quality 50 talks presented at the "International qPCR 2010 Symposium in Vienna" - To get connected visit our web shop => http://eConference.bioeps.com
View our eSeminar and eConference demo videos on http://eseminars.bioeps.com/player/?action=demo
qPCR Symposium USA in November 2010 - http://events.gene-quantification.info/
qPCR Application Workshops - Download course brochure 2010 => http://www.gene-quantification.de/bioeps-course-programm-2010.pdf
Register here => http://site.bioeps.com/index.php?option=com_seminar&Itemid=6

How to do successful gene expression analysis - Siena 20100625

2010-07-19T15:58:00.001+02:00

Check out this SlideShare Presentation:

How to do successful gene expression analysis - Siena 20100625

View more presentations from Biogazelle.

NEW - single-cell qPCR workshop - 18-20 October 2010

2010-07-19T10:47:00.005+02:00

The course focus on experimental setup and data analysis doing RT-qPCR in samples with low nucleic acid amounts and single-cells.
For beginner and advanced qPCR user.
Practical and theoretical parts.
Language: In English.
Location: BioEPS GmbH, Lise-Meitner-Strasse 30, 85354 Freising, Germany (very close to Munich airport MUC)
Link => http://site.bioeps.com/index.php?option=com_seminar&Itemid=6
Day 1:
* Introduction into Methods for single-cell sorting * Single-cell theory and applications * Sampling and cultivation technologies * Basic qPCR theory and applications
Day 2:
* Principles of cDNA synthesis * Pre-amplification * Primer and Probe design * Optimizations and experimental setup using single-cells * Data analysis in single-cell experiments
Day 3:
* Introduction to quantification principles * Absoulte quantification with single-cells * Relative quantification: Gene expression * Calculations using different relative quantification methods * Strategies for normalization of QPCR data
Practical Part:
In the practical part laser microdissection using the Cell-Ector system (MMI) will be presented from tissue material. Interested participants are encouraged to bring their personal sample material.
Furhtermore the AmpliSpeed single-cell RT-PCR system from Advalytix (Beckman Coulter) will be used.

Exiqon at the qPCR Symposium 2010

2010-07-16T09:35:00.002+02:00

Exiqon at the qPCR Symposium 2010
powered by http://www.bioeps.com/
Watch Exiqon’s presentation from the qPCR Symposium 2010 in Vienna. Learn how to overcome the challenges of microRNA profiling in difficult samples.
http://www.exiqon.com/ls/Documents/movies/qPCR-2010/qPCR-2010.html

more real-time qPCR talks are available here http://eseminars.bioeps.com/shop/index.php?page=product&info=6

qPCR Basics eSeminar

2010-07-15T12:45:00.005+02:00

Real-Time PCR: Reference Genes & Data Analysis

2010-07-01T13:19:00.005+02:00

Real-Time PCR: Current Technology and Applications
Publisher: Caister Academic PressEditor: Julie Logan, Kirstin Edwards and Nick Saunders Applied and Functional Genomics, Health Protection Agency, London (2009) ISBN: 978-1-904455-39-4

Chapter 4 - Reference Gene Validation Software for Improved Normalization
J. Vandesompele, M. Kubista and M. W. Pfaffl (2009)
Real-time PCR is the method of choice for expression analysis of a limited number of genes. The measured gene expression variation between subjects is the sum of the true biological variation and several confounding factors resulting in non-specific variation. The purpose of normalization is to remove the non-biological variation as much as possible. Several normalization strategies have been proposed, but the use of one or more reference genes is currently the preferred way of normalization. While these reference genes constitute the best possible normalizers, a major problem is that these genes have no constant expression under all experimental conditions. The experimenter therefore needs to carefully assess whether a certain reference gene is stably expressed in the experimental system under study. This is not trivial and represents a circular problem. Fortunately, several algorithms and freely available software have been developed to address this problem. This chapter aims to provide an overview of the different concepts.
Chapter 5 - Data Analysis Software
M. W. Pfaffl, J. Vandesompele and M. Kubista (2009)
Quantitative real-time RT-PCR (qRT-PCR) is widely and increasingly used in any kind of mRNA quantification, because of its high sensitivity, good reproducibility and wide dynamic quantification range. While qRT-PCR has a tremendous potential for analytical and quantitative applications, a comprehensive understanding of its underlying principles is important. Beside the classical RT-PCR parameters, e.g. primer design, RNA quality, RT and polymerase performances, the fidelity of the quantification process is highly dependent on a valid data analysis. This review will cover all aspects of data acquisition (trueness, reproducibility, and robustness), potentials in data modification and will focus particularly on relative quantification methods. Furthermore useful bioinformatical, biostatical as well as multi-dimensional expression software tools will be presented.

Real-Time PCR: Current Technology and Applications - Book reviews:
"... a comprehensive overview of the RT-PCR technology, which is as up-to-date as a book can be ..." Mareike Viebahn in Current Issues in Molecular Biology (2009)
"... a useful book for students ..." from J. Microbiological Methods
"provides a dual focus by aiming, in the early chapters, to provide both the theory and practicalities of this diverse and superficially simple technology, counter-balancing this in the later chapters with real-world applications, covering infectious diseases, biodefence, molecular haplotyping and food standards." from Microbiology Today
"a reference work that should be found both in university libraries and on the shelves of experienced applications specialists." from Microbiology Today
"a comprehensive guide to real-time PCR technology and its applications" from Food Science and Technology Abstracts (2009) Volume 41 Number 6
"This volume should be of utmost interest to all investigators interested and involved in using RT-PCR ... the RT-PCR protocols covered in this book will be of interest to most, if not all, investigators engaged in research that uses this important technique ... a well balanced book covering the many potential uses of real-time PCR ... valuable for all those interested in RT-PCR." from Doodys reviews (2009)
"provide the novice and the experienced user with guidance on the technology, its instrumentation, and its applications" from SciTech Book News June 2009 p. 64
"... written by international authors expert in specific technical principles and applications ... a useful compendium of basic and advanced applications for laboratory scientists. It is an ideal introductory textbook and will serve as a practical handbook in laboratories where the technology is employed." from Christopher J. McIver, Microbiology Department, Prince of Wales Hospital, New South Wales, Australia writing in Australian J. Med. Sci. 2009. 30(2): 59-60

PCR Optimization Strategies

2010-07-01T13:13:00.000+02:00

PCR Optimization Strategies
RT-PCR Optimization Strategiesfrom Martina Reiter and Michael W. Pfaffl writing in PCR Troubleshooting and Optimization: The Essential Guide PCR technology is based on a simple principle; an enzymatic reaction that increases the amount of nucleic acids initially present in a sample but this powerful method makes it possible to detect specific mRNA transcripts in any biological sample by the application of RT-PCR. The RT-PCR quantitative analysis workflow has several steps, each of which is crucial to the success of the experiment. It starts with a sampling step, followed by nucleic acid extraction and stabilization, cDNA synthesis and finally the qPCR where the mRNA quantification takes place. PCR itself is quite a stable reaction with reproducibility between 2-8% but the number and nature of the pre-PCR steps mean that there are many sources of experimental variance in the workflow. Reliable data can only be produced when the experimental variance is minimized, so the sources of variation must be identified and optimized for each step of each experiment. Typically, however, the pre-PCR steps are neglected and optimization is done for PCR reaction only. Optimization of the whole RT-PCR workflow is important and recommendations to reduce experimental variance and produce more reproducible and reliable results should be followed.
Further reading: PCR Troubleshooting and Optimization: The Essential Guide

The evolution of qPCR

2010-06-29T16:00:00.003+02:00

BioEPS and MMI AG agree on joint Workshops From Single- and Rare Cells to RT-PCR

2010-06-29T12:18:00.002+02:00

BioEPS and MMI AG agree on joint Workshops From Single- and Rare Cells to RT-PCR
BioEPS Munich, an innovative Biotech company, offering services related to molecular technologies, especially real-time qPCR technology and mmi AG Zurich, the leading provider for micromanipulation systems agreed to share competencies.
Both partners provide specific Courses for single cell analysis as of summer 2010 in Zurich and Munich.
For courses, contents and individual offerings contact us at info@molecular-machines.com and info@bioEPS.com

2010-06-28T17:28:00.003+02:00

Check out this SlideShare Presentation about real time PCR:

Real Time Pcr

View more presentations from Nirmala last.

PCR, Real Time PCR

2010-06-28T17:27:00.001+02:00

Check out this SlideShare Presentation:

PCR, Real Time PCR

View more presentations from dineshnbagr.

Next Generation Sequencing & Transcriptome Analysis

2010-06-28T17:24:00.001+02:00

Check out this SlideShare Presentation:

Next Generation Sequencing & Transcriptome Analysis

View more presentations from Bastian Greshake.

MIQE Press Review

2010-06-16T14:47:00.000+02:00

MIQE PRESS REVIEW
http://miqe-press.gene-quantification.info/

Efforts to standardize qPCR data meets mixed reviews
05/25/2010Uduak Grace Thomas - BioTechniques
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Routine lab method's accuracy called into question
Catherine Shaffer - Nature Medicine Vol 16, page 349 (2010)
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The MIQE Guidelines Uncloaked - Speaker: Greg Shipley
8 June 2010 - The MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) guidelines have been presented to serve as a practical guide for authors when publishing experimental data based on real-time qPCR. Each item is presented in tabular form as a checklist within the MIQE manuscript. However, this format has left little room for explanation of precisely what is expected from the items listed and no information on how one might go about assimilating the information requested. This presentation presents an expanded explanation of the guideline items with commentary on how those requirements might be met prior to publication.
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High-Impact Journals, Large Vendors Contributing to Lack of Quality Control for qPCR Data Publication
June 03, 2010 - By Ben Butkus
GÖTEBORG, Sweden – Quality control for publication of quantitative PCR data is still severely lacking – and, in the case of high-profile scientific journals such as Nature and Science, it is "absolutely appalling and scandalous" – according to Stephen Bustin, one of the scientists leading the drive to adopt a set of standards intended to guide high-quality and reproducible qPCR experiments within the field......
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Following MIQE Recommendations - EMBL Heidelberg, Germany
Monday 5 July - Friday 9 July 2010
Since the early descriptions of the use of quantitative Real Time PCR, the technique has been adopted in almost every aspect of life science research and is increasingly used for clinical analysis. Over time protocols and strategies have been tried and tested, amended and developed such that there are currently several different approaches. Protocol variations are evident at each step of the RT-qPCR process, from sample acquisition to data analysis (e.g. sample QC, experimental design, assay design and validation, normalisation, biostatistical interpretation, reporting, etc). It is now apparent that these adaptations may result in differences in the final biological conclusion of the study.
This workshop is based upon the MIQE guidelines. Each step of the RT-qPCR process will be discussed and protocol variations illustrated practically. The student will be instructed in best practice and acceptable alternative strategies.
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A Practical Guide to Publishing RT-qPCR Data That Conform to the MIQE …
In an effort to assist the scientific com- munity in producing consistent, high- quality data from qPCR experiments, the minimum information for publication of quantitative real-time PCR experiments .
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Real-time PCR on SciTopics
UPDATE on 21 January 2010 by Prof Stephen Bustin
Category: Biochemistry, Genetics and Molecular BiologyGuidelines for minimum information required for publication of qPCR data have now been published in Clinical Chemistry
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qPCR Assay Quality Assessment on SciTopics
UPDATE on 21 January 2010 by Prof Stephen Bustin
Category: Biochemistry, Genetics and Molecular BiologyGuidelines for minimum information required for publication of qPCR data have now been published in Clinical Chemistry
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MIQE Guidelines 'Slowly Filtering Through' PCR Community Despite Lack of Journal Enforcement
December 31, 2009 - Although the guidelines are beginning to catch on among researchers and vendors, they appear to have made little or no impact on the quality of the published literature over the last year.
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MIQE PRESS REVIEW
http://miqe-press.gene-quantification.info/

RNA quality control in miRNA expression analysis

2010-06-16T14:44:00.000+02:00

RNA quality control in miRNA expression analysis
Agilent Technologies Application Note 5990 5557 EN
http://tinyurl.com/3aabtf6

RNA quality control in miRNA expression analysis
Christiane Becker, Martina Reiter, Michael W. Pfaffl
Agilent Technologies Application Note 5990-5557EN

It is generally known that total RNA quality has a distinct influence on the validity and reliability of quantitative PCR results. In addition, the recently published MIQE guidelines focus on the pre-PCR steps and state the importance of RNA quality assessment. Various studies showed the impairing effect of ongoing RNA degradation on mRNA expression results. Therefore, the verification of RNA integrity prior to downstream applications like RT-qPCR and mircroarrays is indispensable. A fast and reliable assessment of RNA integrity can be done with the Eukaryote Total RNA Nano Assay of the Agilent 2100 Bioanalyzer. The importance of RNA quality should also be considered in new applications such as the investigation of miRNA expression profiles. With the Agilent Small RNA Assay, Agilent is offering one of the few possibilities for selectively estimating miRNA before expression analysis. However, by now little is known about factors affecting miRNA analysis. Herein, the important impact of total RNA quality on quantification of mRNA and miRNA should be considered.

Now Online - qPCR 2010 eConference

2010-06-10T13:34:00.002+02:00

BioEPS is now presenting seminars and scientific talks online via video stream!

With our new eSeminars and eConference tool a world-wide know-how transfer becomes possible in a fast and simple way.
http://eSeminars.bioeps.com/player/?action=demo
The qPCR 2010 package contains 50 talks from our qPCR Symposium 2010 in Vienna
Topics: MIQE guidelines, High throughput qPCR, RNAi-miRNA, qPCR Data analysis, HRM, Circulating Nucleic Acids, Single Cell qPCR

The complete qPCR 2010 eConference package contains 50 talks in high resolution from the qPCR Symposium 2010 in Vienna with all sessions.

Or you can visit the single sessions including following keynote speakers:

For more information visit our online shop =>

http://eConference.qpcr2010-vienna.net/

REST 2009 software

2010-06-10T13:33:00.000+02:00

REST 2009 Software is a standalone software tool, developed by M. Pfaffl (Technical University Munich) and QIAGEN, for analysis of gene expression data from quantitative real-time PCR experiments. Click the "Resources" tab to download REST 2009 Software free of charge.

Download newest version REST 2009 => http://www.rest.de.com/
Download User Guide => REST 2009 Software User Guide - English (PDF)
REST 2009 Software is intended for molecular biology applications. This software is neither intended for the diagnosis, prevention, or treatment of a disease, nor has it been validated for such use either alone or in combination with other products. Therefore, the performance characteristics of the product for clinical use (i.e., diagnostic, prognostic, therapeutic, or blood banking) are unknown. REST 2009 SoftwareFor gene expression analysis using real-time PCR data from the Rotor-Gene Q and other cyclers
Estimates up and down regulation for gene expression studies
Analysis using randomization and bootstrapping techniques
Graphical data output via whisker-box plots.

Call for Abstracts - qPCR 2010 Event

2009-11-19T17:50:00.000+01:00

qPCR 2010 Event – The ongoing evolution of qPCR

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BioEPS GmbH is organizing the qPCR 2010 Event taking place April 7th – 9th, 2010 in Vienna, Austria. Scientists from all around the world will come to exchange ideas, share experiences, and discuss the exciting future of the perhaps most powerful analytical technology ever developed in the life sciences area – the quantitative real-time polymerase chain reaction (qPCR). We are proud to present 40 scientific talks from invited international scientists and diagnostic companies in the qPCR field who will show their latest research findings and newest technologies. Focus of the event will be - The ongoing evolution of qPCR.

The scientific organization is managed by international well-known scientists in the field of qPCR:

Stephen Bustin Prof. of Molecular Science, School of Medicine, London, UK
Mikael Kubista Prof. of Biotechnology, TATAA Biocenter, Sweden
Christine Mannhalter Prof. for Molecular Diagnostics in Clinical Chemistry, Med. University Vienna, Austria
Jo Vandesompele Prof. at the Center of Medical Genetics, University of Ghent, Belgium
Michael W. Pfaffl Senior Scientist and Reader in Physiology, TUM, Weihenstephan, Germany

Using qPCR the amount of target nucleic acid in a complex sample can be determined with high precision, great accuracy, excellent specificity and the ultimate sensitivity of detecting a single molecule. The technique has revolutionized all molecular sciences and diagnostic applications. Conference presentations will include MIQE guidelines & QM strategies in qPCR, high performance nucleic acid extraction, single-cell applications, Epigenetics & High-Resolution-Melt analysis, circulating nucleic acids, and application involving RNAi and microRNA.
Further developments of qPCR technology will be presented include improved instrumentation, miniaturization, high throughput platforms, cost efficacy, validity, flexibility, quality assessment and reliable Cq calculations, expression data comparisons, and interpretation.
Today there is no field in the life sciences research, molecular biology and diagnostics areas that has not introduced qPCR technology for nucleic acid analysis. The combination with reverse transcription enables determination of mRNA, microRNA and widely opens the window for “Transcriptomics” – the first step of quantitative “Gene Expression Profiling” and “Functional Genomics”.
An Industrial Exhibition will take place parallel to the symposium, with 32 leading biotechnology companies presenting their latest developments, including real-time PCR cyclers, NA extraction robots, consumables, new fluorescence dyes, NA detection and amplification chemistries, as well as real-time PCR data analysis software.

For more information about the qPCR 2010 event contact Dr. Martina Reiter martina.reiter@bioeps.com

Hope to meet you in April in Vienna!

Michael Pfaffl
scientific coordinator

NEW BOOK - The PCR Revolution: Basic Technologies and Applications

2009-11-12T15:38:00.000+01:00

The PCR Revolution: Basic Technologies and Applications
edited by Stephen A. Bustin

The invention of the polymerase chain reaction (PCR) won the Nobel Prize in Chemistry in 1994 and remains one of the most important scientific discoveries of the twentieth century. More than 50,000 researchers in the United States use PCR replication technology, and yet a book has not been published on the subject in more than ten years. In this book, Dr. Stephen A. Bustin, a world-renowned PCR expert, examines in detail the latest innovations and the overall impact of PCR on many areas of molecular research. The book contains personal reflections, opinions, and comments by leading authorities on the many applications of the PCR and how this technology has revolutionized their respective areas of interest. This book conveys the ways in which PCR has overcome many obstacles in life science and clinical research and also charts the PCR's development from time-consuming, low throughput, non-quantitative procedure to today's rapid, high throughput, quantitative super method.

Inside This Book - Browse Sample Pages:
Table of Contents First Pages Index

International qPCR 2010 Symposium & Exhibition in Vienna

2009-10-25T16:37:00.000+01:00

International qPCR 2010 Symposium & Exhibition in Vienna
7th – 9th April 2010
The focus of the qPCR 2010 Event will be “The ongoing evolution of qPCR”

Our online registration and abstract submission tool - CONFTOOL - is available now => http://registration.qpcr2010-vienna.net

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qPCR 2010 in Vienna

International qPCR 2010 Symposium and Exhibition in Vienna 7th – 9th April 2010.
The focus of the qPCR 2010 Event will be “The ongoing evolution of qPCR” representing all new and emerging techniques, applications and data analysis methods. MIQE, HRM, microRNA, CNV, single-cell qPCR, digital PCR, and analysis of circulating nucleic acids will be in the focus of the conference => http://www.qpcr2010-vienna.net/

qPCR Symposium topic - “The ongoing evolution of qPCR”

The symposium focus will be on 40 lectures presented by international recognized experts in their application fields. The emphasis will be on unbiased, didactic information exchange. One third of the talks will be presented by invited speakers, one third of the speakers will be selected from the submitted abstracts and one third will be qPCR company representatives with their newest qPCR technologies.

The focus of the qPCR 2010 Event will be “The ongoing evolution of qPCR” representing all new and emerging techniques, applications and data analysis methods:

Symposium sessions & preliminary titles

MIQE and QM strategies in qPCR
Prof. Stephen Bustin, “The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments”
High throughput quantitative PCR – digital PCR
Prof. Mikael Kubista, “Digital PCR and intracellular expression profiling”
Dr. Philip Day, “High throughput droplet PCR”
Dr. Ken Livak, "title to be announced"
HRM – High Resolution Melting - Epigenetics
Prof. Carl Wittwer, "High Resolution Melting Analysis"
Prof. Claudio Orlando, “High Resolution Melting Analysis in Cancer Diagnosis”
Circulating nucleic acids
Dr. Pamela Pinzani, “Cell free circulating DNA”
Dr. Alfred Schöller, „Targeting the human urine RNAome for tumor diagnostics by qPCR”
Dr. Jim Huggett, "Diagnostic tools for measuring cell free nucleic acids. What can we expect from the next decade?”
Single-cell qPCR
Dr. Michael W. Pfaffl, “Quantitative expression analysis after pre-amp in single WBCs”
Dr. Anders Stahlberg, “Single-cell gene expression
RNAi - microRNA - siRNA Applications – miRNA normalisation
Prof. Jo Vandesompele, “MicroRNA and mRNA gene expression normalization”
Dr. Mirco Castoldi, "Expression profiling of microRNA by quantitative real time PCR, what is available and where to go from there"
qPCR BioStatistics & BioInformatics
Dr. Ales Tichopad, “Statistical aspects of quantitative PCR experiment design and qPCR data analysis”
Dr. Jan Hellemans, “Accurate and objective copy number profiling using real-time quantitative PCR”
Dr. Anders Bergkvist, “Expression profiling - clusters of possibilities”

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Contact information:

Dr. Martina Reiter

BioEPS GmbH
Lise Meitner Strasse 30
85354 Freising Weihenstephan
Germany
qPCR2010@BioEPS.com

Standardization of qPCR and RT-qPCR

2009-08-04T11:02:00.000+02:00

PCR Technology Review: Standardization of qPCR and RT-qPCR
New Guidelines Seek to Promote Accurate Interpretation of Data and Reliable Results
by Stephen A. Bustin, Jo Vandesompele, Michael W. Pfaffl

The perceived ease of use of real-time quantitative PCR (qPCR) and reverse transcription PCR (RT-qPCR) technology has revolutionized life science research. Its effectiveness at amplification and quantification of low levels of nucleic acids has driven the emergence of numerous applications, including cellular mRNA and miRNA quantification, biomarker discovery and validation, microbial quantification, cancer risk assessment, gene dosage determination, and detection of extremely low copy targets for forensic investigations.This, in turn, has resulted in an abundance of publications utilizing qPCR data obtained with diverse reagents, protocols, analysis methods, and reporting formats. Unfortunately, few papers report in detail how these results were obtained. This lack of clarity and transparency has led to concern in the research community over the reliability of qPCR data interpretation and the real danger of the scientific literature being corrupted with publications reporting erroneous and conflicting results.This has already occurred in some cases, resulting, for example, in retraction of a Science “Breakthrough of the Year 2005” report. Now that qPCR has come of age, standardization is needed to ensure its validity, prompting the recent formulation of guidelines to increase experimental transparency, promote consistency between laboratories, and therefore, help assure the publication of valid conclusions.

http://www.genengnews.com/articles/chitem.aspx?aid=2992
http://www.gene-quantification.de/miqe.html#press

MIQE - qPCR Assay Quality Assessment

2009-06-24T14:22:00.000+02:00

The MIQE guidelines: minimum information for publication of quantitativereal-time PCR experiments.
Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT. (PDF)

Clin Chem. 2009 55(4): 611-622
http://www.rdml.org/miqe
http://miqe.gene-quantification.info
MIQE - media & press review

BACKGROUND: Currently, a lack of consensus exists on how best to perform andinterpret quantitative real-time PCR (qPCR) experiments. The problem isexacerbated by a lack of sufficient experimental detail in many publications,which impedes a reader's ability to evaluate critically the quality of theresults presented or to repeat the experiments.
CONTENT: The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelinestarget the reliability of results to help ensure the integrity of the scientific literature, promote consistency between laboratories, and increase experimentaltransparency. MIQE is a set of guidelines that describe the minimum informationnecessary for evaluating qPCR experiments. Included is a checklist to accompanythe initial submission of a manuscript to the publisher. By providing allrelevant experimental conditions and assay characteristics, reviewers can assess the validity of the protocols used. Full disclosure of all reagents, sequences,and analysis methods is necessary to enable other investigators to reproduceresults. MIQE details should be published either in abbreviated form or as anonline supplement.
SUMMARY: Following these guidelines will encourage betterexperimental practice, allowing more reliable and unequivocal interpretation of qPCR results.

qPCR Application workshops

2009-06-19T13:54:00.000+02:00

Still availibility in the JULY qPCR course
27 - 31 Juli 2009 - 3-day qPCR Core Module & 2-day BioStatistics Module

At the TATAA Biocenter Germany, located at BioEPS GmbH, we offer various qPCR application workshops: 3-day qPCR Core Module, 2-day qPCR Biostatistics Module, 3-day microRNA qPCR Module & 3-day single-cell qPCR Module. All courses are held regularly in Göteborg, Sweden, in English and in Freising-Weihenstephan, Germany, in German and English, and in Prague, Czech Republic in English and Czech.
Depending on the occasion the workshop language and the different prices may apply. Further customized workshops and specialized trainings will be held as well across Europe and world-wide.
TATAA Biocenter Germany workshops are powered by BioEPS GmbH, located at the campus of the Technical University of Munich, in Freising-Weihenstephan, very close to the Munich Airport (MUC). For more information and registration, please see our web page => http://tataa.gene-quantification.info/

Course Occasions 2009:
3-day qPCR Core Module (Mon. - Wed.)
2-day BioStatistics & Expression Profiling Module (Thu. - Fri.)
3-day single-cell qPCR Module (Mon. - Wed.)
3-day microRNA qPCR Workshop (Mon. - Wed.)

13 - 15 Juli 2009 (E) NEW microRNA qPCR

27 - 31 Juli 2009 (E) 3-day qPCR Core Module (Mon. - Wed.) & 2-day BioStatistics Module (Thu. - Fri.)

14 - 16 September 2009 (E) NEW single-cell qPCR

19 - 23 October 2009 (E) 3-day microRNA Module (Mon. - Wed.) & 2-day BioStatistics (Thu. - Fri.)

26 - 28 October 2009 (E) 3-day qPCR Core Module (Mon. - Wed.)

16 - 20 November 2009 (E) 3-day microRNA Module (Mon. - Wed.) & 2-day BioStatistics (Thu. - Fri.)

7 - 11 December 2009 (E) 3-day qPCR Core Module (Mon. - Wed.) & 2-day BioStatistics (Thu. - Fri.)

course program spring - summer 2009

course program autumn 2009

A novel and universal method for microRNA RT-qPCR data normalization

2009-06-19T09:35:00.000+02:00

Published in last week's Genome Biology ... ... ...

Jo Vandesompele is senior author on a paper also appearing this week on a new method to normalize miRNA RT-qPCR data. In this study, his team used a mean expression value of all expressed microRNAs in a sample as a normalization factor for miRNA real-time quantitative PCR data and compared it to what's currently used, showing that the mean expression value "outperforms the current normalization strategy in terms of better reduction of technical variation and more accurate appreciation of biological changes," they say in the abstract.

Finally, researchers at the University of Toronto have performed a comparative analysis of metabolism across 193 eukaryotes. Using whole and partial genome and proteome datasets, they found that metabolic enzymes are, in general, "highly conserved," but that certain pathways are more highly conserved than others and that there is a "highly conserved, but nonetheless flexible, 'core' of enzymes largely involved in multiple reactions across different pathways."

Journal reference:

Pieter Mestdagh, Pieter Van Vlierberghe, An De Weer, Daniel Muth, Frank Westermann, Frank Speleman and Jo Vandesompele. A novel and universal method for microRNA RT-qPCR data normalization. Genome Biology, (in press) [link]

qPCR / real-time PCR

qPCR newsletter - January 2012

A MIQE Case Study — Effect of RNA Sample Quality and Reference Gene Stability on Gene Expression Data

Practical Course on Single-Cell Gene Expression Analysis

EMBL Master Course

qPCR newsletter December 2011

GenEx version 5 for FREE download

qPCR NEWS - November 2011 - focus on mirtrons

BioTechniques - Future Methods in PCR

new MIQE APP available on Android Market

Gene-Expression Analysis Exploits More Technologies

qPCR NEWS - October 2011 - focus on MIQE and RefGenes

LIFE SCIENCE TECHNOLOGIES - qPCR Innovations and Blueprints

Next meeting of the New England RNA Data (NERD) club

qPCR NEWS - September 2011 issue

MIQE guidelines Online Translation Service

eConference - qPCR 2011 EVENT is now online available!

qPCR and MIQE Seminar Series

qPCR NEWSLETTER - August 2011 - focus on digital-PCR

NEW MIQE-qPCR APP is online since 3rd August

Go, Go Gadgets - Research gets APP happy

our latest paper - Quantification noise in single cell experiments

more than 1,000 downloads of MIQE_qPCR APP

Follow our qPCR 2011 eConference

Bio-Rad Introduces the First MIQE qPCR App, Now Available at the Apple App Store

Video Tutorial: MIQE and your qPCR data

Evaluierung der qPCR

Follow our new facebook MIQE.qPCR community

MIQE_qPCR APP for iPhone, iPad and iPod

PCR NEWS - February 2011

qPCR 2011 Event Agenda is online

qPCR NEWS - Jan 2011 - Data Analysis & Bioinformatics & qPCR efficiency

VERY VERY COOL - Zheng Lab - Bad Project (Lady Gaga parody)

FINAL CALL for TALK & POSTER submission qPCR 2011 Event

qPCR NEWS Dec 2010 - focus on RNA and DNA quality control for qPCR

An Interview with MIQE Guru and qPCR Expert Stephen Bustin

qPCR 2011 Event - Industrial Exhibition almost fully booked

Next Basic qPCR Application Workshop 17-19 January 2011

Basic qPCR eSeminar

The ongoing evolution of qPCR

qPCR 2011 Event in Freising Weihenstephan

Single-Cell Challenges - GEN article

Join our microRNA qPCR application workshop in November

Making the most of MIQE - MIQE precis

An Interview with MIQE Guru and qPCR Expert Stephen Bustin

The Future of qPCR: Best practices, Standardization, and the MIQE Guidelines

Review: Normalization strategies for microRNA profiling

qPCR NEWS - August 2010 - focus on Copy Number Variations

GenEx offers advanced methods to analyze real-time qPCR data

Real Time QPCR Data Analysis Tutorial (part 2 relative quantification)

Real Time QPCR Data Analysis Tutorial (part 1 absolute quantification)

MIQE Guidelines on baidu.com

qPCR NEWS - July 2010 - focus on PCR efficiency

How to do successful gene expression analysis - Siena 20100625

NEW - single-cell qPCR workshop - 18-20 October 2010

Exiqon at the qPCR Symposium 2010

qPCR Basics eSeminar

Real-Time PCR: Reference Genes & Data Analysis

PCR Optimization Strategies

The evolution of qPCR

BioEPS and MMI AG agree on joint Workshops From Single- and Rare Cells to RT-PCR

PCR, Real Time PCR

Next Generation Sequencing & Transcriptome Analysis

MIQE Press Review

RNA quality control in miRNA expression analysis

Now Online - qPCR 2010 eConference

REST 2009 software

Call for Abstracts - qPCR 2010 Event

NEW BOOK - The PCR Revolution: Basic Technologies and Applications

The PCR Revolution: Basic Technologies and Applicationsedited by Stephen A. Bustin

Inside This Book - Browse Sample Pages:Table of Contents First Pages Index

International qPCR 2010 Symposium & Exhibition in Vienna

Standardization of qPCR and RT-qPCR

MIQE - qPCR Assay Quality Assessment

qPCR Application workshops

A novel and universal method for microRNA RT-qPCR data normalization

The PCR Revolution: Basic Technologies and Applications
edited by Stephen A. Bustin

Inside This Book - Browse Sample Pages:
Table of Contents First Pages Index