title: miRNA expression profiling using Illumina Universal BeadChips.
abstract: We have developed a highly sensitive, specific, and reproducible method for microRNA (miRNA) expression profiling, using BeadArray technology. This method incorporates an enzyme-assisted specificity step, a solid-phase primer extension to distinguish between members of miRNA families. In addition, a universal PCR is used to amplify all targets prior to array hybridization. Using this method, highly reproducible miRNA expression profiles were generated with 100-200 ng total RNA input. The method has a 3.5-4 log (10(5)-10(9) molecules) dynamic range and is able to detect 1.2- to 1.3-fold differences between samples. Expression profiles generated by this method are highly comparable to those obtained with RT-PCR (R (2) = 0.85-0.90) and direct sequencing (R = 0.87-0.89). This method should prove useful for high-throughput expression profiling of miRNAs in large numbers of tissue samples.
authors.affiliation: Illumina, Inc., San Diego, CA, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.forename: Craig S
authors.initials: CS
authors.lastname: April
authors.forename: Jian-Bing
authors.initials: JB
authors.lastname: Fan
mesh_terms: D000818:Animals; D015342:DNA Probes; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D035683:MicroRNAs / Q000032:analysis* / Q000302:isolation & purification; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*
chemical_list: D015342:DNA Probes; D035683:MicroRNAs