miRNA detection probe and miRNA amplification detection method

Abstract

The invention discloses a linear template and a hairpin probe and a method for miRNA amplification detection by using the linear template and hairpin probe. The linear template and hairpin probe disclosed by the invention are easy to design; the mirRNA amplification detection method disclosed by the invention has relatively high amplification efficiency and relatively high detection sensitivity.

Description

technical field [0001] The invention relates to the fields of molecular biology and nucleic acid chemistry, in particular to a miRNA detection hairpin probe and a miRNA amplification detection method. Background technique [0002] MicroRNAs (miRNA) are a class of non-coding RNAs with regulatory functions found in eukaryotic cells, usually about 20-25 nucleotides in length. miRNAs are formed from long hairpin-containing precursors that are cleaved by Dicer enzymes, and then combined with RNA-induced silencing complexes (miRISCs) to recognize corresponding messenger RNAs (mRNAs) through complementary base pairing. Depending on the degree of complementarity, miRNAs can degrade or repress the translation of target mRNAs. Studies have shown that miRNAs are widely expressed in animals and plants and participate in various regulatory pathways, including development, virus defense, hematopoietic process, organ formation, cell proliferation and apoptosis, fat metabolism, etc. miRNA...

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Problems solved by technology

Among them, Northern blot is to perform agarose gel electrophoresis on the RNA sample to be tested under denaturing conditions, and then perform membrane transfer and probe hybridization detection. This method has very high specificity, but the process is complicated and requires a large amount of samples , low sensitivity; the microarray method uses special glass slides or silicon chips to lay thousands or tens of thousands of nucleic acid probes on an area of ​​several square centimeters. Detected by fluorescence or current, this method can realize multiple miRNA analysis at the same time, but the detection cost is high; RT-PCR technology is a technology that combines reverse transcription of RNA and polymerase chain amplification reaction of cDNA. Able to amplify nucleic acids with high efficiency; constant temperature amplification technology can achieve high-efficiency amplification of nucleic acids under constant temperature conditions, in which strand displacement amplification uses polymerase and nickase to cyclically extend and nick specific templates, so It can be amplified to produce a large number of target fragments, but the requirements for equipment are very high

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