Tissue microRNA detection kit based on A11Glo probe fluorogenic quantitative PCR (Polymerase Chain Reaction) and detection method thereof

Abstract

The invention discloses a tissue microRNA detection kit based on A11Glo probe fluorogenic quantitative PCR (Polymerase Chain Reaction) and a detection method adopted by the kit, and relates to microRNA. The kit comprises a box body, a clapboard, a stem-loop reverse transcription reagent bottle and a real-time fluorogenic quantitative PCR reagent bottle. The detection method comprises the following steps: extracting microRNA in a tissue according to an ordinary method, performing reverse transcription on the microRNA into cDNA by using a stem-loop reverse transcription reagent provided by the tissue microRNA detection kit based on the A11Glo probe fluorogenic quantitative PCR, performing real-time fluorogenic quantitative PCR amplification on the cDNA by using a real-time fluorogenic quantitative PCR reagent provided by the tissue microRNA detection kit based on the A11Glo probe fluorogenic quantitative PCR, comprehensively analyzing various data provided by an instrument, setting reasonable thresholds and base lines, and analyzing results.

Description

technical field [0001] The invention relates to microRNA, in particular to a tissue microRNA detection kit based on AllGlo probe fluorescence quantitative PCR and a detection method thereof. Background technique [0002] MicroRNA (miRNA) is a kind of non-coding RNA molecule of about 22 nucleotides widely present in eukaryotic cells, which participates in many physiological and pathological processes in organisms, by regulating gene expression, in cell proliferation, apoptosis , growth and development, cell differentiation, metabolism and other processes play an important role. The specific performance is that miRNA forms the initial primary transcript pri-miRNA to pre-miRNA in the nucleus, and then transports out of the nucleus, forms mature miRNA by shearing, and forms RISC (RNA-induced silencing complex) with Ago protein, etc. Partially inhibit or degrade the target mRNA sequence and participate in the regulation of gene expression (Bartel DP. MicroRNAs: genomics, biogene...

AI Technical Summary

Problems solved by technology

Among them, northern blot technology is one of the early means of RNA detection, but its specificity and sensitivity are low. If the sample RNA content is low or there is degradation, it may not be detected; in situ hybridization technology is used to detect the distribution of miRNA at tissue and cell levels However, the semi-quantitative detection of miRNA at the same time has the disadvantage that it cannot accurately detect low-expression miRNA; microarray chip technology is a high-throughput detection technology that can detect a large number of one or more samples at the same time. miRNA, but there are problems such as time-consuming and labor-intensive chip fabrication, high labeling costs, and low detection sensitivity and specificity

The reverse transcription primer based on the stem-loop structure can specifically recognize and amplify the mature miRNA sequence, but the precursor of the miRNA has not been amplified. The disadvantage of the Taqman-MGB probe for detecting miRNA is that the synthesis is expensive, which is not conducive to popularization.

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