High Throughput Sequencing and Microarray Facility
at Princeton University
Princeton Township, New Jersey, United States of America
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Agilent RNA Microarray
The Agilent RNA microarray technology is an in situ probe generation platform with a fluorescence-based assay to evaluate gene expression by messenger RNA (mRNA) level. An Agilent microarray is created by synthesizing single-stranded 60 nucleotide DNA segments (probes) on a glass slide using a chemical inkjet printing technique (SurePrint). Each probe represents a single gene. Commercial Agilent microarrays include the SurePrint G3 Human Gene Expression microarray and SurePrint G3 Mouse Gene Expression microarray. To create the target sequences for a one-color assay, mRNA is purified from the sample to be analyzed, reverse transcribed, and then amplified by in vitro transcription (IVT) into single-stranded antisense RNA (aRNA). The aRNA is digested and fluorescently labeled. These target sequences are then hybridized to the microarray during which they bind the complementary single-stranded probe and the unbound targets are washed away. The microarray is laser scanned by an Agilent microarray scanner to measure the fluorescent emission, which is correlated to the level of transcription for each gene with Agilent-compatible software. The applications of RNA microarrays include the study of gene regulation, cell fate determination, the identification of disease-related genes, analysis of the effects of drugs on gene expression, and tumor typing for personalized cancer treatments. (Credit: Brooke Anderson-White)
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