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Fluorescence imaging card box and application thereof in nucleic acid aptamer screening

A fluorescent imaging and cartridge technology, applied in the field of medical biochemical detection, can solve the problems of heavy weight, low detection efficiency, lengthy and complicated protein identification steps, etc.

Pending Publication Date: 2021-06-08
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These detection instruments are often large in size and weight, and because of their precision, proteins must be measured in professional laboratories, which makes the steps of protein identification tedious and complicated, and the detection efficiency will be relatively low, which may not meet the requirements of specific experiments. Time requirements, when the laboratory lacks detection equipment, it is even more impossible to complete protein detection

Method used

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  • Fluorescence imaging card box and application thereof in nucleic acid aptamer screening
  • Fluorescence imaging card box and application thereof in nucleic acid aptamer screening
  • Fluorescence imaging card box and application thereof in nucleic acid aptamer screening

Examples

Experimental program
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Effect test

Embodiment 1

[0052] A fluorescent imaging cartridge, comprising a box body 1, the inside of the box body 1 is provided with a bottom plate; wherein, the upper part of the bottom plate is sealed, and a pipetting module and a cleaning solution module are arranged inside; wherein the bottom plate The lower part is open, and there is a reagent storage reaction module.

[0053] Wherein, the height, length and width of the box body 1 are all determined according to the components coupled with it, and its height needs to match the pipetting module and the cleaning solution module, and its length and width match the reagent storage and reaction module.

[0054] Among them, such as figure 1 , figure 2 , Figure 5 and Figure 6 As shown, the core component of the pipetting module is a two-way piece 5, and also includes a vertical pressure plate 2, a first force application rod 31, a second force application rod 32, a first force application point 41, a second force application point 42, two Thr...

Embodiment 2

[0074] Example 2: Application of the fluorescent imaging cartridge described in Example 1 in the screening of nucleic acid aptamers for tumor markers

[0075] (1) Manually add the required reagents to the reagent storage reaction module and the cleaning solution module, that is, the cleaning solution is poured into the cleaning solution storage part 11, the PCR reaction reagent (PCR amplification reagent, a total of 60 μL of forward and reverse primers, 200 μL of Premix, pure 40 μL of water) is poured into the PCR reaction tank 13, and 100 μL of the Cell-SELEX product screening product is poured into the screening product storage tank 12, and the cells are colonized in the culture dish 14; after the upper and lower parts of the cartridge are assembled to ensure sealing;

[0076] (2) Liquid pipetting head 17 draws Cell-SELEX product from screening product storage tank 12, releases to PCR reaction tank 13 again, carries out PCR reaction with PCR reaction reagent;

[0077] (3) Th...

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Abstract

The invention discloses a fluorescence imaging card box and application thereof in nucleic acid aptamer screening. The card box comprises a box body; a bottom plate is arranged in the box body; a pipetting module and a cleaning liquid module are arranged at the upper part of the bottom plate; and a reagent storage reaction module is arranged at the lower part of the bottom plate. According to the portable disposable card box provided by the invention, fluorescent nucleic acid amplification can be achieved in the card box; furthermore, coupling with a fluorescent imaging system of a matched instrument is achieved; and application of a tumour marker screening technology in various environments and conditions is liberated. According to the invention, the tumour marker screening technology is integrated in one nucleic acid aptamer tumour marker screening instrument instead of being only suitable for a laboratory environment; screening operation can be carried out by using the fluorescence card box; various parameters in the screening process are set in advance in control software; after the card box is inserted into the screening instrument, a starting instruction can be given to the screening instrument to perform full-automatic screening; and the experimental skill requirements of a screening instrument detector are also reduced.

Description

technical field [0001] The invention belongs to the field of medical biochemical detection, in particular to a fluorescent imaging cartridge and its application in nucleic acid aptamer screening. Background technique [0002] In recent years, the exponentially enriched ligand system evolution (SELEX) technology, which has been developed rapidly in recent years, uses a large-capacity random single-stranded oligonucleotide library to mix with target substances, and after interacting with target molecules, it screens out compounds that are compatible with target molecules. Specifically binding oligonucleotides, wash away the nucleic acid molecules that are not bound to the target substance, and then separate the nucleic acid molecules that bind to the target substance, and then amplify it in combination with in vitro PCR amplification technology, so that it can be enriched exponentially. Cell-SELEX technology is a method to obtain aptamers that can bind to specific target cells...

Claims

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Application Information

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IPC IPC(8): C12M1/38C12M1/34C12M1/00C12N15/10G01N21/64G01N35/00G01N35/10
CPCC12N15/1048G01N21/6458G01N35/00G01N35/1004G01N35/1009G01N2035/00178C12Q2531/113C12Q2563/107
Inventor 王超刘宾何农跃李智洋赵彦峰陈臻
Owner NANJING MEDICAL UNIV
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