Reagent preparation and storage for amplification of microarray hybridization targets with a fully automated system.
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.
Zhou
Mingjie
M
Rosetta Inpharmatics LLC, Seattle, WA 98109, USA. Mingjie_Zhou@Merck.com
Marlowe
Jon
J
Graves
Jaime
J
Dahl
Jason
J
Riley
Zackery
Z
Tian
Lena
L
Duenwald
Sven
S
Tokiwa
George
G
Fare
Thomas L
TL
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
D018076:DNA, Complementary; D007202:Indicators and Reagents; D018075:RNA, Complementary