The solution and its simple protocol are ideally suited for interrogation of a broad range of small and degraded clinical samples with as little as 500 picograms of input total RNA—a 100-fold reduction from other commercially available in vitro transcription-based (IVT) amplification methods.
At the heart of the solution are NuGEN WT-Ovation™ Systems based on the Single Primer, Isothermal Amplification (SPIA®) technology used to generate cDNA from the most compromised samples—formalin-fixed, paraffin-embedded (FFPE) tissues, laser-captured, micro-dissected (LCM) cells, fine needle aspirates, tissue biopsies, sorted cells, and other sample types that yield small or finite amounts of RNA. The addition of this solution for Agilent Dual-Mode Gene Expression Microarrays completes the list of major expression analysis platforms with which the NuGEN Ovation® family of products is compatible. Others include the Affymetrix GeneChip® microarray platform, the Illumina Genome-Wide Expression BeadChip platform, and qPCR.
A simple, three-step protocol enables researchers to quickly and easily prepare targets in a single day from the most difficult samples:
1. Choose the appropriate whole-transcript amplification system— the WT-Ovation™ FFPE System or the WT-Ovation Pico System.
2. Amplify RNA to generate amplified cDNA.
3. Label amplified cDNA and hybridize to Agilent Dual-Mode Gene Expression Microarrays.
"This is an important breakthrough for the growing number of researchers using Agilent Dual-Mode Gene Expression Microarrays," explained Dr. Joe Don Heath, Senior Director, NuGEN Technical Marketing. "There are valuable repositories of clinical samples around the world that may be key to understanding the causes and progression of cancer, infectious diseases, metabolic and neurological disorders, and more. With this solution, scientists will be able to make significant discoveries using the Agilent gene expression platform, as a precursor to developing targeted diagnostic tools or drug therapies."
David Hirschberg, PhD, of the Human Immune Monitoring Center (HIMC), Stanford University School of Medicine, whose lab has evaluated the new NuGEN protocol, commented, "The sensitive amplification technology offered by NuGEN is changing the questions we are asking, because it enables us to work with extremely small, homogeneous samples that have until now been recalcitrant to global gene expression profiling. The technology is critical in elucidating the intricate biology of the individual components of the immune system and their roles in human health and disease."
The NuGEN Ovation family of RNA amplification and labeling products enables life scientists to conduct sensitive, robust, global gene expression profiling and novel signature discovery. Ovation Systems are platform independent, so researchers may conduct analyses using their preferred gene expression platform.