title: Reagent preparation and storage for amplification of microarray hybridization targets with a fully automated system.
abstract: 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.
authors.affiliation: Rosetta Inpharmatics LLC, Seattle, WA 98109, USA. Mingjie_Zhou@Merck.com
authors.forename: Mingjie
authors.initials: M
authors.lastname: Zhou
authors.forename: Jon
authors.initials: J
authors.lastname: Marlowe
authors.forename: Jaime
authors.initials: J
authors.lastname: Graves
authors.forename: Jason
authors.initials: J
authors.lastname: Dahl
authors.forename: Zackery
authors.initials: Z
authors.lastname: Riley
authors.forename: Lena
authors.initials: L
authors.lastname: Tian
authors.forename: Sven
authors.initials: S
authors.lastname: Duenwald
authors.forename: George
authors.initials: G
authors.lastname: Tokiwa
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Fare
mesh_terms: D001331:Automation; D003365:Costs and Cost Analysis; D018076:DNA, Complementary / Q000032:analysis; D003627:Data Interpretation, Statistical; D006801:Humans; D006824:Hybridization, Genetic; D007202:Indicators and Reagents / Q000737:chemistry*; D019169:Jurkat Cells; D020014:K562 Cells; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics / Q000379:methods*; D018075:RNA, Complementary / Q000032:analysis; D048348:Reverse Transcription
chemical_list: D018076:DNA, Complementary; D007202:Indicators and Reagents; D018075:RNA, Complementary