miRNA microarrays

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MicroRNAs (miRNAs) are small, single stranded RNA molecules encoded in the genomes of plants and animals. These highly conserved, ~20-mer RNAs regulate the expression of genes by binding to the 3'-untranslated regions (3'-UTR) of specific mRNAs with the aid of RISC complex (see figure below). miRNAs have been found to be key regulators in cell growth, cell death, metabolism and cell differentiation. They are becoming increasingly important as more information is discovered in their link to such diseases as cancer, viral disease and neurodevelopment.

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Commercially available miRNA arrays from:

1. Illumina has created two standard fixed MicroRNA expression profiling panels of miRNA assays: one for human and one for mouse. Both panels were developed to assay a complete and broad spectrum of miRNA.

Sample Requirement: Illumina miRNA arrays require 100-200 ng total RNA from cultured cells, fresh frozen, or FFPE tissue

Click here to visit the Illumina Website

2. Agilent has developed a microarray-based application for studying microRNAs (miRNAs) that combines a unique miRNA direct labeling method with their innovative probe design and established high-performance SurePrint inkjet synthesis technology.

Sample Requirement: Agilent miRNA arrays require 100ng total RNA per array (50ng for 2 color arrays). Click here to see our Agilent certified services page.

Click here to visit the Agilent Website

In house printed miRNA arrays using oligos purchased from:

1. Ambion probe set version 2 which is a comprehensive selection of human, mouse, and rat miRNAs from miRBase Sequence Database version 8.0. Probes according to species are represented as follows:

Sample Requirement: The Ambion probe sets are spotted onto aminosaline slides and require two color hybridizations using cy3 and cy5. The Ambion mirVana miRNA isolation kit (cat#AM1562) is highly recommended for small RNA isolations. Subsequently, the purity and quality of the RNA can be determined by 260/280 absorbance ratio of greater than 1.8 and Agilent 2100 BioAnalyzer assay S28/S18 ratio of greater than 1.0 respectively.

Click here to visit the Ambion Website

2. Invitrogen NCode™ Mammalian miRNA Microarray Probe Set Version 1.0 (Cat. # MIRMPS-01) which contains probes targeting all of the known mature miRNAs in mirBase 7.0 for human, mouse, rat, Drosophila, C. elegans, and zebrafish

1. RNA isolation from tissue
2. FlashPage small RNA isolation from total RNA
3. Total RNA Quality Checked with Agilent BioAnalyzer
4. Labeling of RNA using Invitrogen’s NCode miRNA labeling system (MIRLS-20)
5. Hybridization/washing/scanning of arrays using in-house equipment
6. Spot quantification with ImaGene (BioDiscovery).
7. Feature Extraction output files sent to customer (FTP) or in house, custom data analysis.
8. Real time PCR target validation

Please note that if samples do not meet quality control and the customer opts not to continue with the project, a fee of $15/sample will be charged.