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ASSAY and Drug Development Technologies
Reagent Preparation and Storage for Amplification of Microarray Hybridization Targets with a Fully Automated System

To cite this article:
Mingjie Zhou, Jon Marlowe, Jaime Graves, Jason Dahl, Zackery Riley, Lena Tian, Sven Duenwald, George Tokiwa, Thomas L. Fare. ASSAY and Drug Development Technologies. August 2007, 5(4): 551-558. doi:10.1089/adt.2007.060.

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Mingjie Zhou 
Rosetta Inpharmatics LLC, Zymogenetics, Seattle, WA.
Jon Marlowe 
Systems Development, Roche Molecular Diagnostics, Pleasanton, CA.
Jaime Graves 
Rosetta Inpharmatics LLC, Zymogenetics, Seattle, WA.
Jason Dahl 
Rosetta Inpharmatics LLC, Zymogenetics, Seattle, WA.
Zackery Riley 
Methods Development, The Allen Institute, Zymogenetics, Seattle, WA.
Lena Tian 
Department of Molecular & Cellular Discovery, Zymogenetics, Seattle, WA.
Sven Duenwald 
Rosetta Inpharmatics LLC, Zymogenetics, Seattle, WA.
George Tokiwa 
Rosetta Inpharmatics LLC, Zymogenetics, Seattle, WA.
Thomas L. Fare 
Rosetta Inpharmatics LLC, Zymogenetics, Seattle, WA.

The advent of automated systems for gene expression profiling has accentuated the need for the development of convenient and cost-effective methods for reagent preparation. We have developed a method for the preparation and storage of pre-aliquoted cocktail plates that contain all reagents required for amplification of nucleic acid by reverse transcription and in vitro transcription reactions. Plates can be stored at −80°C for at least 1 month and kept in a hotel at 4°C for at least 24 h prior to use. Microarray data quality generated from these pre-aliquoted reagent plates is not statistically different between cRNA amplified with stored cocktails and cRNA amplified with freshly prepared cocktails. Deployment of pre-aliquoted, stored cocktail plates in a fully automated system not only increases the throughput of amplifying cRNA targets from thousands of RNA samples, but could also considerably reduce reagent costs and potentially improve process robustness.

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