Quantitative end-point RT-PCR gene,,,expression measurement using the,,,Agilent 2100 Bioanalyzer and,,,standardized RT-PCR
This application note describes how the Agilent 2100 bioanalyzer can
be used in combination with Standardized RT (StaRT)-PCR reagents
from Gene Express National Enterprises (G.E.N.E.), Inc. (Huntsville,
AL, USA) to obtain highly reproducible, standardized gene expression
measurement with RT-PCR. The internal standards used in StaRT-PCR
control for variable loading of PCR products into electrophoresis channel,
and allow quantitative gene expression measurement at the endpoint
of PCR, without real-time analysis. Gene expression measurement
of 15 genes was more reproducible with the Agilent 2100 bioanalyzer
than with the ABI Prism 310 Genetic Analyzer or agarose gel electrophoresis
methods. The mean coefficient of variance (CV) for these
three methods was 0.26, 0.39 and 0.50, respectively.
We combined the Agilent 2100 bioanalyzer
with a quantitative
method for standardizing gene
expression, Standardized RT
(StaRT)-PCR (Willey et al, Am. J.
Resp. Cell Mol. Biol., 19, 6, 1998),
to produce a sensitive, reproducible
method for quantitative
end-point PCR measurement of
gene expression. The Agilent 2100
system markedly simplifies electrophoresis.
There is no need to
prepare agarose gels and, following
results are provided by PCR product
quantification software. The
internal standards used in StaRTPCR
control for variable loading
of PCR products into electrophoresis
channels, and allow
quantitative gene expression measurement
at the end point of PCR,
without real-time analysis. StaRTPCR
used with agarose gel electrophoresis
has been validated in
independent studies (Loitsch et al,
Clinical Chemistry, 45, 619, 1999;
Allen et al, Am. J. Resp. Cell Mol.
Biol., 21, 693, 1999; Mollerup et al,
Cancer Research, 59, 3317, 1999).
Source:Page: All 1 2 3 4 5 6 7 Related biology technology :1
. Mx4000 Multiplex Quantitative PCR System*,PPP2
. Improved Quantitative Selectivity of Clenbuterol in Human Urine using
High Resolution on the TSQ Quantum Mass Spectrometer3
. The LightCycler is the only System for Quantitative PCR that is as Flexible
. Quantitative Measurement of Cell Proliferation
Using the BrdU ELISA: A Comparison
Between Colorimetric and Chemiluminescent Detection5
. Detection of Genetically Modified Soybean in Processed Foods Using Real-Time Quantitative PCR with SYBR Green I Dye on the DNA Engine Opticon 2 System6
. Qualitative and Quantitative Assessment of Plasmid DNA Isolation, Rev A7
. Quantitative Analysis of Protein-DNA Associations in Vivo Using Real-Time PCR, Rev A8
. Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A9
. Quantitative Analysis of GFP in Plant Extracts Using the VersaFluor Fluorometer System10
. Multiplexed Quantitative Peptide Assays for Protein Biomarkers of Cardiovascular Disease in Human Plasma11
. Quantitative Analysis of the Cell Cycle in Imaging Applications
(Date:7/10/2014)... , July 10, 2014 Research ... addition of the "International Photonic Integrated Circuit ... Forecasts to 2019" report to their offering. ... The concept of photonic integration traces its ... century. The promise of photonic integration went unexplored ...
(Date:7/10/2014)... to the International Atomic Energy Agency (IAEA) the ... acquiring sufficient quantities of plutonium or highly enriched ... device. The IAEA also notes that most cases ... which can be challenging to detect with most ... appearing this week in the Journal of ...
(Date:7/10/2014)... WASHINGTON D.C., June 17, 2014 Using microscopic ... of specific gases, researchers at MIT,s Quantum Photonics ... detection levels in the parts-per-billion range. Optical sensors ... to their high signal-to-noise ratio, compact, lightweight nature, ... optical gas sensors had been developed before, the ...
(Date:7/10/2014)... Understanding the basis of psychiatric disorders has been extremely ... increase risk but are insufficient to cause disease. Now ... Cell Press journal Cell Stem Cell describe ... genetic risks interact with other risk factors or environmental ... Their research pinpoints a genetic variant that may predispose ... Breaking Biology Technology:International Photonic Integrated Circuit (Monolithic Integration, Hybrid Integration, Module Integration) Market - Forecasts to 2019 2Sensitive detection method may help impede illicit nuclear trafficking 2Swell new sensors 2New strategy could uncover genes at the root of psychiatric illnesses 2
Mouse GM CSF Construction Kit provides antigen affinity purified polyclonal capture and tracer antibodies, and antigen standard for development of approximately ten microplate assays....
... Inventory Storage Racks for Standard 3" ... fit all makes and models of ... of high quality stainless steel and ... stainless steel or plastic boxes. ...Biology Products: