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Reagent preparation and storage for amplification of microarray hybridization targets with a fully automated system.

Zhou M, Marlowe J, Graves J, Dahl J, Riley Z, Tian L, Duenwald S, Tokiwa G, Fare TL

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 degrees C for at least 1 month and kept in a hotel at 4 degrees 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.

Published 4 September 2007 in Assay Drug Dev Technol, 5(4): 551-8.
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