qPCR NEWS - May 2010 - focus on transcriptional noise in single-cells

Editor www.Gene-Quantification.info via methods%40net.bio.net (by editor from gene-quantification.info)
Fri May 28 07:13:10 EST 2010


qPCR NEWS - May 2010 - focus on transcriptional noise in single-cells





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  -  single-cell qPCR sub-domain with focus on transcriptional
noise  =3D>  http://singlecell.gene-quantification.info
- UPDATE  -  MIQE press & media review  =3D>  http://MIQE-PRESS.gene-quanti=
fication.info
- International qPCR 2010 symposia  =3D>  http://EVENTS.gene-quantification=
.info
- qPCR Application Workshops  -  and more ... ... ...


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single-cell qPCR Single-cell molecular-biology is a relatively new
scientific branch in biology. The first single-cell analysis were
involved in the characterization of mitochondrial DNA in 1988. Single-
cell DNA analysis, in particular genomic DNA, is important and may be
informative in the analysis of genetics of cell clonality, genetic
anticipation and single-cell DNA polymorphisms. Nowadays for most
scientists the quantitative transcriptomics in a single-cell is much
more important, and the analytical method of choice is the
quantitative real-time RT-PCR. The relative abundance of single mRNAs
and their up- or down-regulation in a single cell, compared to their
neighbour cells, is the goal. The need for quantitative single-cell
mRNA analysis is evident given the vast cellular heterogeneity of all
tissue cells and the inability of conventional RNA methods, like
northern blotting, RNAse protection assay or classical block RT-PCR,
to distinguish individual cellular contributions to mRNA abundance
differences.

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The focus of the updated papers in on transcriptional noise in single
cells:

http://singlecell.gene-quantification.info/

- Stochastic gene expression in a single cell
- Nature, nurture, or chance: stochastic gene expression and its
consequences
- Noise in Gene Expression: Origins, Consequences, and Control
- Use of high throughput sequencing to observe genome dynamics at a
single cell level
- Transcriptional noise and cellular heterogeneity in mammalian
macrophages
- Stochastic gene expression during cell differentiation: order from
disorder?
- Resolving cell population heterogeneity: real-time PCR for
simultaneous multiplexed gene detection in multiple single-cell
samples
- Circulating tumour cells in clinical practice: Methods of detection
and possible characterization
- Quantitative transcription factor analysis of undifferentiated
single human embryonic stem cells
- Single cell analysis of transcriptional activation dynamics
- Single-cell gene expression profiling using reverse transcription
quantitative real-time PCR
- Spatial expression profiles in the Xenopus laevis oocytes measured
with qPCR tomography
- Quantitative RT-PCR gene expression analysis of laser microdissected
tissue samples
- Laser capture microdissection and single-cell RT-PCR without RNA
purification

http://www.gene-quantification.de/singlecell3.html

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MIQE - media & press review

Efforts to standardize qPCR data meets mixed reviews
05/25/2010   Uduak Grace Thomas;  BioTechniques

One year ago, an international team of researchers proposed guidelines
for the publication of real-time PCR experiments. Since then, there
has been mixed response from the scientific community. Uduak Thomas
investigates the reasons behind the resistance.
Read more =3D> http://MIQE-press.gene-quantification.info


Routine lab method's accuracy called into question
Catherine Shaffer
Nature Medicine Vol 16, page 349 (2010)

In 2002, four years after first sparking public controversy over
whether the measles, mumps and rubella vaccine causes autism, Andrew
Wakefield reported a possible molecular mechanism for the connection.
He claimed that a form of irritable bowel disease, which he called
autistic enterocolitis, was triggered by the measles virus (Molec.
Pathol. 55, 84=9690, 2002). That finding, however, was based on a
=93defective experimental technique,=94 Stephen Bustin, a molecular
biologist at Barts and the London School of Medicine and Dentistry,
told a US federal court in 2007. The problem: Wakefield had
incorrectly applied the common laboratory protocol known as
quantitative real-time polymerase chain reaction (qPCR) to come to his
conclusions.
Read more =3D> http://MIQE-press.gene-quantification.info


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. More events =3D> http://events.gene-quan=
tification.info



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qPCR 2010 in Vienna
   International qPCR Symposium & Exhibition
   7-9th April 2010
   http://www.qpcr2010-vienna.net/
   Topic:   "The ongoing evolution of qPCR"


330 attendee from 37 contries and 39 international companies in the
industrial exhibition participate at the event 7 - 9th April 2010 in
Vienna.

During the symposium the presented talks and slides were recorded in
high sound quality and movie resolution and they will be made
available as eConfernce qPCR 2010 on our streaming server
eConference.qPCR2010-Vienna.net  or   http://eConference.bioeps.com

Up to now you can listen to our eSeminar and eConference promotion
trailer  =3D>  http://trailer.qPCR2010-Vienna.net
All symposium participants have FREE access for a limted time to the
recorded talks and to the updated online proceedings including all
participants contact data.

If you are interested to listen to the talks and presented slides you
are welcome to register as USER in our ConfTool registration system
We will inform upon the eConference is available!
The talks by the plenary lecturer, selected invited academic and
industrial speakers will be published in METHODS special qPCR Vol 50
issue 4  (April 2010)  with the title =93The ongoing evolution of
qPCR=94   =3D> TOC  and  HTML and PDFs


Symposium sessions:

   MIQE and QM strategies in qPCR
   High throughput quantitative PCR =96 digital PCR
   HRM =96 High Resolution Melting - Epigenetics
   CNA - Circulating nucleic acids
   Single-cell qPCR
   RNAi - microRNA - siRNA Applications =96 miRNA normalisation
   qPCR BioStatistics & BioInformatics


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BioEPS GmbH - qPCR Application Workshops

Life Science is still a growing sector and new methods and
technologies are continously developed. Therefore permanent training
and education becomes so important.

With our specific course program we are offering a range of high-
quality course modules, in cooperation with different companies to
give a general and independent overview of existing qPCR technologies
and systems. Our course issues are based on skilled know-how from own
research studies and publications.

Our aim is to point out a critical way of thinking to increase the
quality and outcome of experimental data.


All courses are held regularly in Freising-Weihenstephan, Germany, in
German and English language.
Further customized workshops and specialized trainings will be held as
well across Europe and world-wide.
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 =3D>
http://workshops.gene-quantification.info

Course Occasions 2010:

3-day qPCR Basic Module
2-day BioStatistics & Expression Profiling Module
3-day single-cell qPCR
2-day microRNA qPCR
1-day HRM
2-das qPCR-R data analysis   NEW !
1-day Project Management   NEW !
2-day Quality Management  NEW !


Course dates 2010:

14 - 15  June 2010 (E)   2-day microRNA & qPCR (Mon.-Tue.)
12 - 14  July 2010  (E)   3-day qPCR Basic Module (Mon. - Wed.)
15  July 2010  (E)   1-day HRM Module (Thu.)
4 - 6  October 2010  (E)   3-day qPCR Basic Module (Mon. - Wed.)
7  October 2010  (E)    1-day HRM Module (Thu.)
11 - 12 October  2010  (E)   2-day qPC-R - data analysis using R
packages (Mon.-Tue.)
18 - 20 October  2010  (E)   3-day single-cell & qPCR (Mon. - Wed.)
21 - 22 October  2010  (E)   2-day BioStatistics Module (Thu. - Fri.)
8 - 9  November 2010  (E)   2-day microRNA & qPCR (Mon.-Tue.)
29 November - 1 December 2010  (E)   3-day qPCR Basic Module (Mon. -
Wed.)
2 - 3 December 2010  (E)   2-day BioStatistics Module (Thu. - Fri.)
(E)

Download course brochure 2010 =3D> http://www.gene-quantification.de/bioeps=
-course-programm-2010.pdf

Register here =3D> http://site.bioeps.com/index.php?option=3Dcom_seminar&It=
emid=3D6


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Forward Please send the qPCR NEWS to further scientists and friends
who are interested in qPCR !


Best regards,

Michael W. Pfaffl
responsible Editor of the Gene Quantification Pages
http://www.gene-quantification.info



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