Researchers from Applied Biosystems and the University of Ghent Demonstrate Effectiveness of Megaplex(TM) miRNA Profiling Tools on Small Samples
Study Describes Simultaneous Profiling of 450 Human MicroRNAs From
Samples as Small as a Single Cell
FOSTER CITY, Calif.--(BUSINESS WIRE)--Oct. 29, 2008--Researchers
from the Ghent University Hospital in Belgium and Applied Biosystems
Inc. (NYSE:ABI) have published a study demonstrating the effectiveness
of a new method for profiling microRNAs in small samples. The study,
entitled "High-throughput stem-loop RT-PCR miRNA expression profiling
using minute amounts of input RNA," describes the simultaneous
profiling of 450 human microRNAs from samples as small as a single
cell. The paper appears online in the journal Nucleic Acids Research.
The study provides validation for a new workflow for miRNA analysis
developed by Applied Biosystems to enable highly sensitive,
high-throughput detection of miRNAs from extremely small biological
MicroRNAs are a recently discovered class of non-coding small RNA
molecules that are receiving attention from the scientific community
for their important role in regulating gene expression and for recent
research implicating these molecules in complex disease processes such
as cancer. Since the discovery of miRNAs, more than 800 human miRNAs
have been identified and many more are awaiting experimental
validation. In order to harness the potential of this important class
of gene regulators, scientists require high-throughput and sensitive
detection methods capable of measuring miRNA expression from very
small biological samples, including single cells.
The purpose of the study was to test the ability of new tools
based on Applied Biosystems' stem-looped, primer-based real-time PCR
approach and industry-standard TaqMan(R) chemistry to accurately
identify miRNAs in three different neuroblastoma cancer cell lines,
both with and without a pre-amplification step. In the study,
sensitivity to single-cell levels was achieved using a novel approach
incorporating an early-access version of the company's recently
commercialized Megaplex(TM) Primer Pools.
Previous studies have reported the use of pre-amplification in
combination with the stem-loop procedure; however, no published study
thus far has evaluated the effects of the pre-amplification step on
the fidelity of miRNA expression measurement. For that reason, one of
the main goals of the study was to evaluate whether pre-amplification
would compromise the accuracy of the results.
The researchers in this study reported unbiased and efficient
pre-amplification efficiency, irrespective of miRNA expression level,
using this method. In fact, pre-amplification actually enabled the
detection of miRNAs that were undetectable in the experiments without
pre-amplification, despite the use of 40-fold less starting material.
The Megaplex Primer Pools consist of the Megaplex RT Primers and
the Megaplex PreAmp Primers. The Megaplex(TM) RT Primers are a set of
highly multiplexed reverse transcription (RT) primers that greatly
reduce the number of RT reactions required, thereby streamlining the
workflow and reducing the sample input required to create a
comprehensive miRNA profile. Megaplex(TM) PreAmp Primers, used in
conjunction with the TaqMan(R) PreAmp Master Mix, enable amplification
of the starting RNA sample prior to performing detection using
real-time PCR, significantly increasing sensitivity when working with
"The high specificity and sensitivity of this method makes it
ideal for high-throughput miRNA expression profiling," said Jo
Vandesompele, Ph.D., professor at Ghent University Hospital. "This
novel technology from Applied Biosystems opens the way for profiling
the miRNAome from small cell populations or individual cells."
Other technologies enable profiling of multiple miRNAs in a single
experiment, but such approaches generally require significant amounts
of input RNA - 100ng or more - and therefore preclude the use of very
small clinical biopsies or analysis of small subsets of cells or
single cells. This limits researchers' abilities to conduct important
experiments for diseases such as cancer in which often only minute
samples are collected, or originate from archived formalin fixed
paraffin embedded samples in which the genetic material may be
degraded and RNA yield is relatively low.
"The ability to detect and accurately quantify all known miRNAs
from a few cells or even a single cell with these new tools enables
researchers to look further and deeper into the important role of
miRNAs in living systems," said Caifu Chen, Ph.D., scientific fellow
at Applied Biosystems. "We expect that researchers will now be able to
identify novel biomarkers for complex diseases such as cancer, as well
as better understand how miRNAs regulate development of a single
neuron or embryo."
The Megaplex miRNA tools used in this study are now commercially
available from Applied Biosystems for use with the TaqMan(R) MicroRNA
Arrays and with individual TaqMan(R) MicroRNA Assays. Together, these
tools are expected to streamline miRNA analysis workflows for life
science researchers and provide comprehensive coverage of miRNA
expression in small biological samples or single cells.
The study is available online at: http://nar.oxfordjournals.org.
Applied Biosystems is a global leader in the development and
commercialization of instrument-based systems, consumables, software,
and services for the life-science market. Combined with miRNA
isolation kits and analysis products from its Ambion business, the
company offers one of the life-science industry's most comprehensive
lines of microRNA analysis tools. The miRNA TaqMan Assays, the miRNA
TaqMan Arrays, along with a complete line of TaqMan Array Gene
Signature Panels, and more than 1,000,000 individual TaqMan(R) Gene
Expression Assays are available on the Applied Biosystems website.
More information about Applied Biosystems' complete line of gene
expression analysis solutions is available at
www.appliedbiosystems.com or at www.allgenes.com.
About Applied Biosystems Inc.
Applied Biosystems Inc. (formerly known as Applera Corporation) is
a global leader in the development and marketing of instrument-based
systems, consumables, software, and services for academic research,
the life science industry and commercial markets. Driven by its
employees' belief in the power of science to improve the human
condition, the company commercializes innovative technology solutions
for DNA, RNA, protein and small molecule analysis. Customers across
the disciplines of academic and clinical research, pharmaceutical
research and manufacturing, forensic DNA analysis, and agricultural
biotechnology use the company's tools and services to accelerate
scientific discovery, improve processes related to drug discovery and
development, detect potentially pathogenic microorganisms, and
identify individuals based on DNA sources. Applied Biosystems has a
comprehensive service and field applications support team for a global
installed base of high-performance genetic and protein analysis
solutions. Applied Biosystems Inc. is headquartered in Norwalk, CT. On
June 12, 2008, Applera Corporation and Invitrogen Corporation (NASDAQ:
IVGN) announced that their Boards of Directors had approved a
definitive merger agreement under which Invitrogen will acquire all of
the outstanding shares of Applied Biosystems stock. The merger is
subject to customary closing conditions and is targeted to close in
mid-November 2008, subject to receipt of antitrust clearance under the
European Council Merger Regulation. Further information regarding the
merger is contained in a joint proxy statement/prospectus and
supplement that have been mailed to stockholders of the company and
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available at http://www.appliedbiosystems.com. All information in this
news release is as of the date of the release, and Applied Biosystems
does not undertake any duty to update this information unless required
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