DAPI redyeing solution and preparation method and application thereof

A technology of counterstaining solution and solution, which is applied in the field of DAPI counterstaining solution and its preparation, and can solve problems such as low signal-to-noise ratio and difficult clinical interpretation

Pending Publication Date: 2021-05-18
河南赛诺特生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the nucleoplasm ratio of tumor cells is larger than that of normal cells, and in order to observe whether the fluorescent signal is in the nucleus, DAPI can help determine the location of cell signals and distinguish tumor cells. There will be a low signal-to-noise ratio, making clinical interpretation difficult

Method used

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  • DAPI redyeing solution and preparation method and application thereof
  • DAPI redyeing solution and preparation method and application thereof
  • DAPI redyeing solution and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0027] Embodiment 1DAPI counterstain solution 1 and preparation method thereof

[0028] This embodiment provides DAPI counterstaining solution 1, said DAPI counterstaining solution A is mainly composed of 10 mg of p-phenylenediamine, 100 μL of PBS buffer solution with pH=9.0, 0.1 μL of TritonX-100, 100 μL of formamide, DAPI 250ng, glycerol 790uL composition.

[0029] The preparation method of DAPI counterstain solution 1 comprises the steps:

[0030] 1) Prepare p-phenylenediamine solution, 1% TritonX-100 solution and DAPI stock solution respectively;

[0031] 2) Mix the ingredients in step 1) with formamide and glycerin to obtain the product.

[0032] Wherein, the p-phenylenediamine solution in step 1) is prepared by dissolving the PBS buffer solution with pH=9.0: weigh 10 mg of p-phenylenediamine and dissolve it in 1 ml of PBS solution;

[0033] Preparation of PBS buffer solution with pH=9.0: Weigh 8g of NaCl, 0.2g of KCl, 3.63g of Na2HPO4·12H2O, 0.24g of KH2PO4, dissolve ...

Embodiment 2D

[0036] Embodiment 2DAPI counterstain solution 2 and preparation method thereof

[0037] This embodiment provides DAPI counterstain solution 2, and the DAPI counterstain solution A is mainly composed of p-phenylenediamine 10 mg, pH=9.0 PBS buffer 100 μL, TritonX-100 1.0 μL, formamide 100 μL, DAPI 250ng, glycerol 700uL composition.

[0038] The preparation method of DAPI counterstain solution 2 comprises the steps:

[0039] 1) Prepare p-phenylenediamine solution, 1% TritonX-100 solution and DAPI stock solution respectively;

[0040] 2) Mix the ingredients in step 1) with formamide and glycerin to obtain the product.

[0041] Wherein, the p-phenylenediamine solution in step 1) is prepared by dissolving the PBS buffer solution with pH=9.0: weigh 10 mg of p-phenylenediamine and dissolve it in 1 ml of PBS solution;

[0042] Preparation of PBS buffer solution with pH=9.0: Weigh 8g of NaCl, 0.2g of KCl, 3.63g of Na2HPO4·12H2O, 0.24g of KH2PO4, dissolve them in 900ml of double-disti...

Embodiment 3D

[0045] Embodiment 3DAPI counterstain solution 3 and preparation method thereof

[0046] This embodiment provides DAPI counterstain solution 3, said DAPI counterstain solution A is mainly composed of p-phenylenediamine 10 mg, pH=9.0 PBS buffer 100 μL, TritonX-100 1.5 μL, formamide 100 μL, DAPI 250ng, glycerol 650uL composition.

[0047] The preparation method of DAPI counterstain solution 3 comprises the steps:

[0048] 1) Prepare p-phenylenediamine solution, 1% TritonX-100 solution and DAPI stock solution respectively;

[0049] 2) Mix the ingredients in step 1) with formamide and glycerin to obtain the product.

[0050] Wherein, the p-phenylenediamine solution in step 1) is prepared by dissolving the PBS buffer solution with pH=9.0: weigh 10 mg of p-phenylenediamine and dissolve it in 1 ml of PBS solution;

[0051]Preparation of PBS buffer solution with pH=9.0: Weigh 8g of NaCl, 0.2g of KCl, 3.63g of Na2HPO4·12H2O, 0.24g of KH2PO4, dissolve them in 900ml of double-distilled...

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Abstract

The invention relates to a DAPI redyeing solution and a preparation method and application thereof. The DAPI redyeing solution is mainly prepared from p-phenylenediamine, Triton X-100, formamide, DAPI and glycerinum. Every 1mL of the DAPI redyeing solution is prepared from 10mg of p-phenylenediamine, 100 microliters of PBS buffer solution with pH of 9.0, 0.1-1.5 microliters of TritonX-100, 10-150 microliters of formamide, 250ng of DAPI, and 600-800 microliters of glycerinum. An existing DAPI redyeing solution is improved, the signal background is reduced by adding 1% Triton-X-100 and formamide in a certain proportion, the signal-to-noise ratio is increased, and clinical interpretation is convenient. The DAPI redyeing solution is applied to fluorescence in-situ hybridization, a more accurate experiment result can be obtained, and a more reliable diagnosis basis is provided for clinicians.

Description

technical field [0001] The invention belongs to the technical field of molecular pathological diagnosis, and in particular relates to a DAPI counterstain solution and its preparation method and application. Background technique [0002] Fluorescence in situ hybridization (FISH) is a non-radioactive in situ hybridization technology developed on the basis of the original radioactive in situ hybridization technology in the late 1980s. Its principle is based on the DNA base pair Complementarity, after hybridizing the fluorescently labeled single-stranded DNA probe with the DNA in the sample to be tested, observe the position of the fluorescent signal on the chromosome through a fluorescence microscope to reflect the situation of the corresponding chromosome and gene, and realize the detection of the DNA in the cell and tissue samples Qualitative and quantitative analysis of chromosomes or genes. FISH technology has the characteristics of simple operation, fast detection, easy o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6841
CPCC12Q1/6841C12Q2563/107
Inventor 胡娟李三华齐华李贵喜李艳敏王少辉霍清园张绍敏刘文弟刘玲玲
Owner 河南赛诺特生物技术有限公司
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