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Method for detecting ERGIC3 mRNA based on DNA molecules

A technology of DNA molecules and detection probes, which is applied in the field of detection of target ERGIC3mRNA based on DNA molecules, and can solve problems such as inability to detect ERGIC3mRNA

Active Publication Date: 2021-07-27
ZUNYI MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] Existing methods cannot detect ERGIC3 mRNA quickly and effectively

Method used

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  • Method for detecting ERGIC3 mRNA based on DNA molecules
  • Method for detecting ERGIC3 mRNA based on DNA molecules
  • Method for detecting ERGIC3 mRNA based on DNA molecules

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Experimental program
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Embodiment 1

[0048] Such as figure 1 It is a schematic diagram of the method for detecting ERGIC3 mRNA based on DNA molecules of the present invention.

[0049] (1) Feasibility verification results of nucleic acid testing:

[0050] Agarose gel (3%) electrophoresis to verify the ortho-catalyzed hairpin self-assembly reaction, the results are as follows figure 1 shown. Lane 8 is a 500bp DNA marker (Takara), used as a molecular weight reference control for electrophoresis. Lanes 5, 6 and 7 are the corresponding bands of H1, H2 probes and ERGIC3 alone. Lane 4 is the electrophoresis band of the ERGIC3+H1 reaction system, and a lagging band relative to lanes 6 and 7 can be seen, which is due to the assembly of ERGIC3+H1 to form a large molecular weight DNA hybrid, which leads to a slowdown in the electrophoretic movement rate. But from lane 3, it can be seen that ERGIC3 and H2 cannot form DNA hybrids, that is, they do not react. Lane 2 is the electrophoresis band of the assembled chain ERGI...

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Abstract

The invention discloses a method for detecting ERGIC3 mRNA based on DNA molecules, which belongs to the technical field of genes. The method comprises the steps of aiming at target ERGIC3 mRNA, carrying out programmed design and synthesis on detection probes H1 and H2 as recognition elements and a substrate probe sub as a report element; transferring 2.5 mu L of a 2 mu M probe H1, 2.5 mu L of a 2 mu M probe H2 and 5 mu L of a 2 mu M substrate probe sub into an amplification system, adding ERGIC3 mRNA with the concentration of 10 <-14 > to 10 <-8 > into an amplification detection system, performing the whole reaction system in 50 mu L of 1 * Nebuffer 2, and incubating at 37 DEG C for 1 hour; then carrying out fluorescence detection by using an M2 spectrum with the excitation wavelength of 645 nanometers and recording the emission wavelength range of 655 to 725 nanometers. The detection method provided by the invention can realize simple, rapid, sensitive and specific detection of the target ERGIC3 mRNA.

Description

technical field [0001] The invention relates to gene molecular detection technology, in particular to a method for detecting target ERGIC3 mRNA based on DNA molecules. Background technique [0002] ERGIC3 (endoplasmic retinal-Golgi intermediate compartment 3) is a type II transmembrane protein localized in the endoplasmic retinal and Golgi bodies. The main function is to participate in the early transport of proteins from the endoplasmic reticulum to the Golgi apparatus and the glycosylation of glycosidases. Studies have found that ERGIC3 is a new lung cancer marker, which may show new application prospects for early diagnosis and treatment monitoring of lung cancer. [0003] Traditional methods for analyzing mRNA levels mainly include Northern blot, microarray and real-time polymerase chain reaction. Among them, Northern blot and microarray techniques are hindered in practical application due to lack of sensitivity and multi-step operation. Quantitative PCR is of great h...

Claims

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Application Information

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IPC IPC(8): C12Q1/6844
CPCC12Q1/6844C12Q2531/119C12Q2563/107C12Q2521/345
Inventor 吴明松许永杰翟薇孙瑜萍陈晓易朱琴琴刁波徐玺耀
Owner ZUNYI MEDICAL UNIVERSITY
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