qPCR NEWS - January 2008 - Special Topic: High Resolution Melt
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:
- HRM - High Resolution Melt
- updates on qPCR INFOPORTAL
- qPCR application workshops in 2008
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High Resolution Melting (HRM)
HRM is a novel, homogeneous, close-tube, post-PCR method, enabling
genomic researchers to analyze genetic variations (SNPs, mutations,
methylations) in PCR amplicons. It goes beyond the power of classical
melting curve analysis by allowing to study the thermal denaturation
of a double-stranded DNA in much more detail and with much higher
information yield than ever before. HRM characterizes nucleic acid
samples based on their disassociation (melting) behavior. Samples can
be discriminated according to their sequence, length, GC content or
strand complementarity. Even single base changes such as SNPs (single
nucleotide polymorphisms) can be readily identified. The most
important High Resolution Melting application is gene scanning - the
search for the presence of unknown variations in PCR amplicons prior
to or as an alternative to sequencing. Mutations in PCR products are
detectable by High Resolution Melting because they change the shape of
DNA melting curves. A combination of new-generation DNA dyes, high-end
instrumentation and sophisticated analysis software allows to detect
these changes and to derive information about the underlying sequence
constellation.
http://HRM.gene-
HRM Applications
The introduction of HRM has renewed interest in the utility of DNA
melting for a wide range of uses, including:
- Mutation discovery (gene scanning)
- Screening for loss of heterozygosity
- DNA fingerprinting
- SNP genotyping
- Characterization of haplotype blocks
- DNA methylation analysis
- DNA mapping
- Species identification
- Somatic acquired mutation ratios
- HLA compatibility typing
- Association (case/control) studies
- Allelic prevalence in a population
- Identification of candidate predisposition genes
With HRM, these and other applications are done using low-cost generic
dyes where previously custom labeled probes such as TaqMan® or
fluorescence resonance energy transfer (FRET) probes were required.
HRM is thus a simpler and much more cost-effective way to characterize
samples.
------------
http://HRM.gene-
- Introduction
- Workflow
- HRM assay design manual
- Mutation Scanning by High Resolution Melt Analysis:
Evaluation of Rotor-Gene 6000, HR-1 and 384-well LightScanner
- High Resolution Melts - TALKs
- HRM Dyes: LC Green SYTO9 Eva Green BEBO
- Genotyping with PCR - How to choose the right approach (by Tevik
Dorak in The Scientist)
- High-Resolution Melting Curve Analysis Novel Strategies For
PCR-Based Mutation Scanning and
Genotyping (by Michael Hoffmann, Karine Raymond and Jochen Hurlebaus)
- Assay: High-Resolution Melting & Unlabeled Probes (by GEN)
- Mutation Scanning of the Cytidine Deaminase Gene by High-Resolution
Melting Curve Analysis
Using the LightCycler 480 System (by Roche Applied Science in
Bionity.com)
- HRM (by LTF Labortechnik)
- HRM application papers
- Further HRM publications by Roche Applied Science & Corbett Life
Science
------------
With the new qPCR INFO PORTAL and all the presented tools we will help
you with to find the right information about qPCR and related topics
in Molecular Biology in the literature and in the World Wide Web.
=> Papers / Protocols / Methods / Databases / Alets / Feeds / Books /
Forums / E-mail / Directory
http://infoportal.
------------
Upcoming Events World-wide academic and commercial qPCR Events
http://events.
Symposia, Meetings, Conferences, Workshops, Seminars, Online-Seminars,
qPCR Education Program, ...etc...
Please submit your qPCR event here => events@gene-
------------
WORKSHOP
TATAA Biocenter Germany - qPCR Application workshops
At the TATAA Biocenter Germany we offer qPCR application workshops,
the 3-day Core Module and a 2-day Biostatistics Module. qPCR courses
are held in regularly in Göteborg, Sweden, in English and in
Freising-Weihenstep
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 courses are held in cooperation with the Institute
of Physiology, located at the Technical University of Munich, in
Freising-Weihenstep
(MUC). For more information and to register for the qPCR application
workshops, please see our web page:
http://tataa.
Course Occasions 2008:
3-day qPCR Core Module (Mon. - Wed.) and 2-day BioStatistics
Module (Thu. - Fri.)
* 3rd - 7th March 2008 (in Freising, Germany, English language)
* 5 - 9th May 2008 (in Freising, Germany, Kurs wird in DEUTSCH
gehalten, German language)
* 7 - 11th July 2008 (in Freising, Germany, English language)
Please register here => http://www.tataa.
------------
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-
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