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Telomere probe and preparation method and application thereof

A telomere and probe technology, applied in the field of telomere probes and their preparation, can solve problems such as complex detection steps and inaccurate detection, and achieve high probe quantum yield, improved resolution, and good photostability Effect

Active Publication Date: 2018-02-23
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Purpose of the invention: In order to overcome the problems of complex detection steps and inaccurate detection encountered in telomere detection, the present invention provides this kind of telomere probe to realize accurate detection of telomere

Method used

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  • Telomere probe and preparation method and application thereof
  • Telomere probe and preparation method and application thereof

Examples

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Embodiment 1

[0034]Example 1: Take 0.5mL of the oil phase CdSSe / ZnS quantum dot solution (the concentration of the oil phase CdSSe / ZnS quantum dot solution is 5mg / mL), dissolve it in 1.5mL of absolute ethanol, shake it evenly, and centrifuge at 8000round / min 20min, centrifuge once. Remove the supernatant, add 2ml of chloroform, 100μL of mercaptopropionic acid, stir evenly, then add 200μL of tetramethylammonium hydroxide, adjust the pH of the solution so that the pH of the solution is ≥ 11, stir vigorously for 1.5-2h, then add 2mL of Deionized water, after stirring, let it stand for stratification, take the upper layer, add three times the volume of absolute ethanol to the reaction solution, centrifuge at 8000rmp / min for 20min, centrifuge once, wash the obtained precipitate with absolute ethanol once, and obtain the aqueous phase CdSSe / ZnS quantum dots, disperse 2.5mg of the aqueous phase CdSSe / ZnS quantum dots precipitate prepared above in 2mL of water to obtain an aqueous phase CdSSe / ZnS...

Embodiment 2

[0035] Embodiment 2: Take 0.5mL of oil phase CdSSe / ZnS quantum dot solution (concentration of oil phase CdSSe / ZnS quantum dot solution is 5mg / mL), dissolve in 1.5mL of absolute ethanol, shake evenly, and centrifuge at 8000round / min 20min, centrifuge once. Remove the supernatant, add 2ml of chloroform, 500μL of mercaptopropionic acid, stir well, then add 200μL of tetramethylammonium hydroxide, adjust the pH of the solution so that the pH of the solution is ≥ 11, stir vigorously for 1.5-2h, then add 2mL of Deionized water, after stirring, let it stand for stratification, take the upper layer, add three times the volume of absolute ethanol to the reaction solution, centrifuge at 8000rmp, and after 20min, wash the obtained precipitate with absolute ethanol once to obtain the aqueous phase CdSSe / ZnS For quantum dots, 2.5 mg of the aqueous phase CdSSe / ZnS quantum dot precipitate prepared above was dispersed in 2 mL of water to obtain an aqueous phase CdSSe / ZnS quantum dot solution, ...

Embodiment 3

[0036] Embodiment 3: Take 0.5mL of oil phase CdSSe / ZnS quantum dot solution (concentration of oil phase CdSSe / ZnS quantum dot solution is 5mg / mL), dissolve in 1.5mL of absolute ethanol, shake evenly, and centrifuge at 8000round / min 20min, centrifuge once. Remove the supernatant, add 2ml of chloroform, 300μL of mercaptopropionic acid, stir well, then add 200μL of tetramethylammonium hydroxide, adjust the pH of the solution to make the pH ≥ 11, stir vigorously for 1.5-2h, then add 2mL of Ionized water, after stirring, let it stand for stratification, take the upper layer, add three times the volume of absolute ethanol to the reaction solution, centrifuge at a speed of 8000 round / min for 20min, and centrifuge once, and wash the obtained precipitate once with absolute ethanol to obtain the aqueous phase CdSSe / For ZnS quantum dots, 2.5 mg of the aqueous phase CdSSe / ZnS quantum dot precipitate prepared above was dispersed in 2 mL of water to obtain an aqueous phase CdSSe / ZnS quantu...

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Abstract

The invention discloses a telomere probe and a preparation method and application thereof. According to the telomere probe, a single stranded DNA is coupled onto the surface of the CdSSe / ZnS quantum dots, the telomere probe can be combined with the cell telomere by in-situ hybridization technique, and the telomere hybridized with the telomere probe is imaged by a single molecular positioning technique to realize the detection of the telomere. The telomere probe realizes the specific marking of the telomere, has higher resolution due to the adoption of the single molecule positioning microscopeimaging, and overcomes the limitation by the optical diffraction limit of the fluorescence microscopic technique. The telomere probe and the preparation method and the application thereof can be usedfor further study of the telomere structure.

Description

technical field [0001] The invention relates to a DNA probe and its preparation method and application, in particular to a telomere probe and its preparation method and application. Background technique [0002] Telomere is a special structure at the end of eukaryotic chromosome, and its sequence is a tandem repeat sequence rich in G nucleotides. The ends of chromosomal DNA are swollen into granules that cover the ends of each chromatid like a cap, so each pair of chromosomes has 2 chromatids and 4 telomeres. There are many differences between this structure and the ends formed by chromosome breaks, such as the inability to fuse with each other, and the cell can only replicate when the structure of the chromosome ends is intact. In vertebrates, the telomeric sequence is a double-stranded structure composed of TTAGGG repeat sequence and its complementary sequence, and one of the single-stranded 3'overhangs forms an overhang structure consisting of hundreds of bases . This ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6841C12N15/11C12N15/10
CPCC12Q1/6841C12Q2563/107C12Q2543/10C12Q2521/327C12Q2521/537
Inventor 宗慎飞王乐陈晨王著元崔一平
Owner SOUTHEAST UNIV
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