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This chapter describes how to create a multiplexed miRNA profiling assay using readily available technology and reagents for midplex miRNA profiles (midplex used to denote mutliplexing up to about 100 miRNAs per well). The assay described is a direct hybridization assay, i.e., it involves no amplification. Further, the reagents are simple and an assay can be assembled in a few hours with simple design rules and very simple execution. The assay execution takes less than 5 h and involves only liquid addition, which makes it possible to create a multiplexed assay for a large number of miRNAs and run the entire profile for hundreds of samples in a matter of a couple of days. In addition, the profiles can be created for any short RNAs, i.e., they are not tied to human or any other species database for miRNAs, and the described assay works for both animal and plant miRNAs. |
