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Kit and method for detecting microRNA (ribonucleic acid)-122

A kit and asymmetric technology applied in the field of kits for detecting microRNA-122

Inactive Publication Date: 2013-10-23
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no rapid, specific and simple kit and method for detecting miR-122

Method used

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  • Kit and method for detecting microRNA (ribonucleic acid)-122
  • Kit and method for detecting microRNA (ribonucleic acid)-122
  • Kit and method for detecting microRNA (ribonucleic acid)-122

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Embodiment 1

[0028] Embodiment 1, hairpin probe and kit for detecting microRNA-122

[0029] 1. Selection of detection target sequences and design of asymmetric hairpin probes

[0030] (1) Select the target sequence for detection according to the molecular biological characteristics of miRNA.

[0031] through literature or Genbank databases ( http: / / www.ncbi.nlm.nih.gov / ) to seize the human miRNA sequence related to cancer and determine it as the detection target sequence.

[0032] (2) Design of specific asymmetric hairpin probes

[0033] Asymmetric hairpin probes were designed for the target sequence using Array Designer 4.0 software, and the designed candidate probe sequences were submitted to the Genbank database for specificity analysis. Submit the designed asymmetric hairpin probes to The DINAMelt Web Server (http: / / mfold.rna.albany.edu / / ?q=DINAMelt / Hybrid2) for secondary structure analysis and between probes, between probes and The complementarity analysis of the target sequence...

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Abstract

The invention discloses a kit and method for detecting microRNA (ribonucleic acid)-122. The kit comprises asymmetrical hairpin probes shown in SEQ ID NO.1 and SEQ ID NO.2. Compared with traditional RT-PCR (reverse transcription-polymerase chain reaction), northern blot and other technologies, the kit has the prominent advantages that (1) the kit carries out liquid-phase hybridization reaction, is closer to the natural state of molecular reaction, solves the problem of high steric hindrance of solid-phase hybridization, and improves the hybridization reaction efficiency; (2) the kit has high detection efficiency and high detection speed, and the traditional nucleic acid detection technology usually needs a few hours and even a dozen hours while the technology carries out molecular recognition and cascade signal amplification simultaneously, can complete nucleic acid detection within ten minutes and does not need to carry out RT-PCR on samples in advance; and (3) the reaction system is simple, dispenses with enzyme reaction and is low in cost.

Description

technical field [0001] The invention relates to the field of biology, in particular to a kit and a method for detecting microRNA-122. Background technique [0002] microRNAs are a class of non-coding single-stranded small molecule RNAs found in a variety of eukaryotic cells and viruses on endogenous chromosomes, with a length of 21-25 nt, which are highly conserved in evolution and can pass through their targets. Complementation of the 3' untranslated region (3'UTR) of the mRNA molecule leads to inhibition of translation of the mRNA molecule. miRNA is involved in regulating the expression of more than one-third of eukaryotic genes, inhibiting the translation of target genes, causing mRNA degradation, etc., and has very important biological functions. Further studies have shown that miRNAs play key roles in cell proliferation, differentiation and apoptosis, neuronal polarity, insulin secretion, brain morphogenesis, heart formation, embryonic development, and individual death...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 夏涵梁盼盼黄庆府伟灵
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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