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Quantitative
Gene Expression Analysis |
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Company Profile
SEQUENOM is committed to providing the best genetic analysis products
that translate genomic science into superior solutions for molecular
medicine and biomedical research. The cornerstone of SEQUENOM's
approach is the Company's proprietary MassARRAY® system, a
high-performance DNA analysis platform, which efficiently and precisely
measures
the amount of genetic target material and variations. The
system is able to deliver reliable and specific data from complex
biological samples and from trace amounts of genetic target material.
Contact
Info
sales@sequenom.com
www.sequenom.com
CORPORATE HEADQUARTERS
3595 John Hopkins Court, San Diego, CA 92121-1331
Phone (858) 202-9000 Fax (858) 202-9001
EUROPEAN OFFICE
Mendelssohnstrasse 15D, D-22761, Hamburg, Germany
Phone (+49) 40-899676-0 Fax (+49) 40-899676-10
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Quantitative
Gene Expression Analysis Overview
MassARRAY QGE
from Sequenom combines Competitive PCR with MALDI-TOF mass spectrometry
(MS) analysis to offer
a highly accurate, sensitive, and high-throughput method for the
quantitative analysis of gene expression in multiple samples. MassARRAY
QGE is scalable, allowing you to perform assays without compromising,
accuracy, sensitivity, or reproducibility, thus solving the
“multi-plate replication gap” seen with other quantitative gene
expression technologies. In addition, the MassARRAY method does not use
chemiluminescence, fluorescence or other secondary-labeling approaches,
allowing unambiguous multiplexing and the inclusion of multiple
normalization standards in each assay.
MassARRAY
QGE is the method of choice over real-time PCR for validation of
microarray data, for the investigation of coding and noncoding
transcripts – and whenever sensitivity and accuracy are required in
transcription analysis.
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- Instrumentation The foundation of the
MassARRAY system is the MassARRAY MALDI-TOF mass spectrometer. Several
system options are available depending on throughput requirements.
Sequenom developed the MassARRAY MALDI-TOF MS Analyzer systems to
specifically meet the requirements of moderate to high throughput
quantitative genetic analysis. Accurate analysis is ensured by the
configuration of the MassARRAY MS optics and the 384-pad SpectroCHIP.
The mass of each primer extension product is resolved using the
Time-of-Flight (TOF) tube on the MassARRAY MALDI-TOF MS. The separated
masses are analyzed in the MS using MassARRAY QGE software.
- Software The accuracy,
sensitivity and automation of the MassARRAY QGE system are enabled and
optimized by the MassARRAY software. The software includes
AssayDesigner for PCR and hME primer and competitor oligonucleotide
design and Typer™ software for data analysis, management and reporting
functions. The QGE software package contains full data analysis and
management capabilities built on an Oracle data management system.
- Reagents The
MassARRAY QGE method uses robust, familiar biochemistry: Competitive
PCR and the hME primer extension reaction. The oligonucleotides used in
these assays are
unlabeled synthetic oligonucleotides that can
be ordered from your own supplier or through Assays-by-Sequenom. In
addition, Sequenom can serve as a your source for process-tested
enzymes and reagents, or you can continue to use your current trusted
suppliers for quality reagents. (Check with us for process approved
reagents and enzymes.)
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MassARRAY QGE data collection and analysis
is fully software supported. The user interface
can be formatted by each user to display different views pertaining to
data analysis. Experiment data is exportable as a text file for
analysis in third party gene expression software.
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MassARRAY QGE quantifies cDNA over a wide concentration range with a
low RSD, independent of concentration. |
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Download Application Notes
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Features and
Benefits |
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FEATURES
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BENEFITS
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Competitive
PCR
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| • Internal standard in every assay |
• High precision, CV <3%
• Wide dynamic range; nM to aM
• Relative and absolute quantitation capable
• Short amplicons enable use of trace and degraded
• RNA samples
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Separate
PCR and Analysis Steps
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• End-reaction assay methodology
• Quantitation independent of PCR kinetics
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• Highest sensitivity – measures single molecule range
• Sensitive down to 1 – 10 cell range
• Measure as low as 10% change in relative gene expression
• Automated for higher throughput
• Multiple assay plates can be standardized and normalized |
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Mass
Spectrometry Analysis
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• TOF tube
provides HPLC-like product separation
• for every assay
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• Single assay multiplex genes of interest and normalizing
• HKG genes
• All genes of interest can be analyzed – any organism
• Uses unlabeled oligonucleotides
• Multiple plate technology – ideal for microarray validation |
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Flexible
Assay Design
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• Complete user control
• AssayDesigner software
• Validated assays through Assays-by-Sequenom option
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• Allele-specific gene expression analysis
• Analyze transcription of individual members of multigene
• families
• Analyze alternate splice-sites
• Take advantage of exon-exon junction sequence
• information for mRNA specificity |
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