METHOD FOR DETECTING PRIMARY AND PRECURSOR MIRNAS

The invention relates to a method for detecting primary and precursor miRNAs, which comprises the following steps of positioning a target miRNA, designing primers for the target miRNA and detecting RT-QPCR. Specifically: locate the target miRNA according to one or more of transcription start site, CpG island, expressed sequence tags, polyA, Cap analysis of gene expression and CIS-PET/Ditags; when design the primers, the setting range of the forward primer is from the 5'-terminal of the sequence to the GU motif, and the setting range of the reverse primer is from the CNNC motif to the 3'-terminal; and the Tm values of the forward primer and the reverse primer are 55~61oC, and the maximum Tm difference between the forward primer and the reverse primer is 2oC; perform reverse transcription and PCR on the sample, and establish a coordinate system of the fluorescence intensity and the target miRNA to realize the detection of the target miRNA. According to the annotation of the upstream and downstream boundaries of primary and precursor miRNA, the quantitative amplification primers are designed according to their transcription features, and the design method is simple and feasible, and has good potential application value for miRNA markers with clinical diagnostic significance.