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DNA hybridization enhancement solution as well as preparation method and application thereof

A technology of enhancement solution and buffer solution, which is applied in the field of preparation of DNA hybridization enhancement solution, and can solve the problems of prolonging hybridization time by formamide

Pending Publication Date: 2021-04-09
BEIJING SEARCH BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although the use of the alterant reduces TM, formamide significantly prolongs the hybridization time

Method used

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  • DNA hybridization enhancement solution as well as preparation method and application thereof
  • DNA hybridization enhancement solution as well as preparation method and application thereof
  • DNA hybridization enhancement solution as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Paraffin section samples were prepared as follows:

[0053]1. The thickness of the paraffin section is 4-5μm, and the slice is picked up with a special glass slide for in situ hybridization, and the slice is baked at 65°C; the slice is sequentially immersed in xylene for 10min×2, absolute ethanol for 5min×2, and air-dried for 2-5min;

[0054] 2. Microwave heating, put the Coplin bottle into the microwave oven and heat at 100% power until the solution starts to boil. When the solution starts to boil, immediately heat it in a microwave oven at 10-20% power for 15 minutes to keep the solution at a temperature slightly below the boiling point. Immediately move into purified water at room temperature and soak for 3 minutes;

[0055] 3. Submerge the slide in 37°C preheated protease working solution (purchased from Sigma, catalog number 77160-100G) and incubate for 20 minutes. Observe under the microscope that the cell outline is clear and evenly dispersed, then transfer it t...

Embodiment 2

[0064] Cytology samples were prepared as follows:

[0065] 1. Harvest cells: collect the cultured or uncultured cells into a graduated centrifuge tube, and discard the supernatant by centrifugation;

[0066] 2. Hypotonicity: Add 0.075mol / L KCl solution pre-warmed at 37°C and gently blow the cell mass with a straw to mix well, then place in a constant temperature water bath at 37°C for hypotonic treatment for 20-30 minutes;

[0067] 3. Pre-fixation: add the newly prepared fixative solution (methanol: glacial acetic acid = 3:1), carefully blow and mix with a pipette, and centrifuge to discard the supernatant. Fixation: Add fixative, mix well by pipetting, and fix for 20 minutes. The supernatant was discarded by centrifugation. Re-fixation: Repeat step 4 2 times or more until the cell pellet is washed white and clean.

[0068] 4. Preparation of cell suspension: add an appropriate amount of freshly prepared fixative according to the number of cells, and pipette the cells to mak...

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Abstract

The invention discloses a DNA hybridization enhancement solution as well as a preparation method and application thereof. The DNA hybridization enhancement solution comprises components with the following concentrations: a solution of a buffer agent as a solute; a hybridization promoter: 2-60 g / 100 ml of a polymer, 10-80% of an organic solvent in volume percentage, and 0.5-10 g / 100 ml of protein in mass volume percentage; and salt: 1 mM to 750 mM. The preparation method comprises the following steps: adding the hybridization promoter into the buffer solution and mixing, and finally adding the salt and mixing to obtain the DNA hybridization enhancement solution. The DNA hybridization enhancement solution is applied to a DNAFISH product. The DNA hybridization enhancement solution shortens hybridization time, has a good hybridization effect, has high specificity and sensitivity, and reduces an amount of probes to one-fifth of the original amount and saves cost of probes.

Description

technical field [0001] The present invention relates to the molecular analysis field of in situ hybridization, relates to a DNA hybridization enhancement solution and its preparation method and application, in particular to a DNA hybridization enhancement solution and a DNA hybridization enhancement solution which can be used in combination with a DNA molecular probe Preparation. Background technique [0002] In situ hybridization (Insituhybridization, ISH) is a technology produced by the combination of molecular biology, histochemistry and cytology. [0003] In situ hybridization technology can be divided into genome in situ hybridization technology, fluorescence in situ hybridization technology, multi-color fluorescence in situ hybridization technology and in situ PCR. [0004] Fluorescence in situ hybridization (FISH) is a non-radioactive molecular cytogenetic technique developed on the basis of radioactive in situ hybridization in the late 1980s. A new in situ hybridiz...

Claims

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Application Information

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IPC IPC(8): C12Q1/6832C12Q1/6841
CPCC12Q1/6832C12Q1/6841
Inventor 杨德刚高云周赵伟张荣刘明坤叶锋
Owner BEIJING SEARCH BIOTECH
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