Life Technologies Launches High-Throughput Solution for Global Screening of Human miRNAs
by Mauricio Minotta - 04/12/11
CARLSBAD, Calif. – April 12, 2011 – Life Technologies Corporation today announced the introduction of its TaqMan® OpenArray® MicroRNA Panels, a next-generation profiling approach designed to address an industry need for high-throughput technology that facilitates global screening using more than 750 of the company’s pre-designed human miRNAs assays.
Research has shown that dysregulation of miRNAs, single stranded molecules that serve as global regulators of gene expression, has been directly linked to cancer, immune disorders and heart disease, among other conditions. However, the short nature of miRNA sequences (~22 nucleotides) makes it challenging to adapt traditional DNA-based analysis tools for miRNA detection and quantification. Current methods for global profiling of miRNA expression with the required high-throughput capabilities are limited and complicated.
Life Technologies addresses these hurdles with a two-pronged approach: combining for the first time its high-throughput OpenArray® Real-Time PCR Platform with its market-leading TaqMan® MicroRNA Assays. The dual approach enables a single researcher to generate more than 27,000 highly sensitive and accurate miRNA data points in a working day by screening 754 unique human miRNAs per sample for up to 36 samples using a simple workflow (three samples per OpenArray® slide, three slides per instrument run, and four runs within an 8-hour period).
“We are pleased with the high throughput and ease of use of the OpenArray platform for miRNA TaqMan assays,” said Victor Ambros, Ph.D., Co-director, RNA Therapeutics Institute Silverman, Professor of Natural Sciences Program in Molecular Medicine at the University of Massachusetts. “We also find attractive the lower costs per sample on account of the reduced use of reagents compared to other real-time PCR platforms.”
Unlike Life Technologies’ conventional TaqMan® Assay products, miRNA assays use a novel stem-loop primer during the reverse transcription (RT) reaction. Stem-loop primers provide several advantages. First, the RT product extends the miRNA sequence creating a longer cDNA molecule that is more amenable to detection using standard TaqMan chemistry.
Their double-stranded structure sterically interferes with hybridization of the RT primer to longer miRNA precursors and genomic DNA. Finally, base stacking of the stem enhances stability of miRNA RT primer complexes improving efficiency and ultimately assay sensitivity.
Technical challenges associated with high-density qPCR analysis, such as changes in PCR stoichiometry during thermal cycling and the need for stringent fluid isolation between adjacent reaction containers to prevent cross contamination, are addressed with the OpenArray® Platform, technology and intellectual property Life Technologies acquired through its acquisition of BioTrove in 2009.
The high-throughput OpenArray® gene expression and genotyping analysis system is centered on a stainless steel nanofluidic array the size of a microscope slide. Each array is photolithographically patterned and etched to form a rectilinear array of 3,072 through-holes arranged in 48 groups of 64 holes.
A series of vapor and liquid deposition steps covalently attach a PCR compatible polyethylene glycol (PEG) hydrophilic layer to the interior surface of each through-hole, and a hydrophobic fluoroalkyl layer to the exterior surface of the array. The differential hydrophilic-hydrophobic coating facilitates precise loading and isolated retention of PCR reagents and sample in each through-hole.
“By leveraging the accuracy and high-throughput capabilities of our TaqMan and OpenArray technologies, we’re providing researchers with highest confidence miRNA screening results using the simplest workflow,” said Sam Raha, Vice President and General Manager of NextGen qPCR & Content for Life Technologies. “Through this approach, what would have taken weeks can now be accomplished in days.”
About Life Technologies (www.lifetechnologies.com)
Life Technologies Corporation is a global biotechnology company dedicated to improving the human condition. Our systems, consumables and services enable researchers to accelerate scientific and medical advancements that make life even better. Life Technologies customers do their work across the biological spectrum, working to advance the fields of discovery and translational research, molecular medicine, stem cell-based therapies, food safety and animal health, and 21st century forensics. The company manufactures both molecular diagnostic and research use only products. Life Technologies' industry-leading brands are found in nearly every life sciences lab in the world and include innovative instrument systems under the Applied Biosystems and Ion Torrent names, as well as, the broadest range of reagents with its Invitrogen, GIBCO, Ambion, Molecular Probes and TaqMan products. Life Technologies had sales of $3.6 billion in 2010, has a workforce of approximately 11,000 people, has a presence in approximately 160 countries, and possesses one of the largest intellectual property estates in the life sciences industry, with approximately 3,900 patents and exclusive licenses. For more information on how we are making a difference, please visit our website: http://www.lifetechnologies.com.
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