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µParaflo RNA Microarray
The µParaflo RNA microarray technology from Atactic Technologies and LC Sciences is a microfluidic, digital light synthesized platform to evaluate gene expression by messenger RNA (mRNA) level in small samples. A uParaflow array is created by synthesizing single-stranded DNA segments (probes) in a microfluidic chamber using a digital photolithography technique. Each probe represents a single gene. Several target sequence generating methods are compatible with uParaflo. In general, 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). Fluorescently labeled nucleotides are incorporated during IVT and the labeled aRNA is digested. These target sequences are then hybridized to the microarray during which they bind the complementary single-stranded probes. The unbound targets are then washed away and the microarray is laser scanned by a uParaflo-compatible instrument to measure the fluorescent emission. The emission is correlated to the level of transcription for each gene with uParaflo-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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