Principle

In the proprietary miRNA plate assay, one miRNA molecule is flanked by a capture oligo and a biotinated detection oligo through two bridges oligos. One of the bridge oligos is partially hybridized with the miRNA molecule and the capture oligo and another with the miRNA and the detection oligo. The hybrid is immobilized onto plate through hybridization with an immobilized oligo and detected by a streptavidin-HRP conjugate and chemiluminecscent substrate. This hybrid structure is sensitive to the sequence of the miRNA molecule. One nucleotide difference will prevent the formation of the hybrid and therefore miRNA isoform can be differentiated, which normally is hard to tackle with Northern blot. In addition, the sensitivity of the assay is higher than miRNA Northern blot assay.