Kit for rapidly detecting telomerase activity and application of kit

A technology of telomerase and kit, applied in the direction of microbial measurement/testing, fluorescence/phosphorescence, biochemical equipment and methods, etc., can solve the problems of not being widely used, not having advantages, and complicated steps, etc. Scale and potential for commercial application, avoiding false positive or false negative results, simple steps

Inactive Publication Date: 2012-09-26
云南路易斯中药现代化工程技术研究中心
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  • Abstract
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AI Technical Summary

Problems solved by technology

Although this method is widely used, it still has its shortcomings that cannot be ignored: the steps are relatively complicated, time-consuming, and it is easy to get false positive or false negative results, etc.

Method used

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  • Kit for rapidly detecting telomerase activity and application of kit
  • Kit for rapidly detecting telomerase activity and application of kit
  • Kit for rapidly detecting telomerase activity and application of kit

Examples

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Example Embodiment

[0036] Example 1.

[0037] A kit for rapid detection of telomerase activity, which is characterized by comprising 5 and 4 reagent tubes; wherein the first reagent tube 1 contains the extension reaction mixture, and the second reagent tube 2 contains the labeled molecular beacon sequence , The third reagent tube 3 contains ExoIII, and the fourth reagent tube 4 contains 1x NEB Buffer 1.

[0038] The application of a kit for rapid detection of telomerase activity is implemented as follows:

[0039] 1. Extraction of telomerase from cell samples

[0040] Collect tumor cells (HeLa cell line, purchased from ATCC) and human embryonic lung diploid fibroblasts (WI-38 cell line, purchased from ATCC), and centrifuge the cells to 200 μL / 10 6 Resuspend the ratio of cells in the lysis buffer (1x CHAPS Lysis buffer, ATCC) and let stand on ice for 30 min; then centrifuge at 12000 g at 4°C for 20 min; collect the supernatant and store at -80°C to obtain two types Granzyme supernatant, namely HeLa cell...

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Abstract

The invention relates to a kit for rapidly detecting the telomerase activity, and the kit is characterized by comprising a kit body and four reagent tubes, wherein the reagent tubes are respectively filled with an extension mixture, a marked molecular beacon sequence, ExoIII, and 1xnebbuffer1. Instrument consumables required by the kit disclosed by the invention are low in cost, and the kit has commercial potential in the large-scale detection of telomerase activity.

Description

Background technique [0001] Telomere (Telomere) is a DNA repeating sequence located at the end of chromosomes, which can maintain the integrity of chromosomes. In 1990, Harley et al. confirmed that the length of telomeres can gradually shorten as the number of cell divisions increases. This finding establishes a strong link between telomeres and cellular aging. The length of the telomeric repeats may act as a kind of molecular clock. The life span of somatic cells of people of different ages is obviously different, and the length of their telomeres is also different. is shortened with age. Somatic cells from newborns can be subcultured for 80-90 generations in vitro, while somatic cells from 70-year-olds can only be subcultured for 20-30 generations in vitro, and the repeat length of telomeres is also much shorter. [0002] Telomerase (Telomerase) is an enzyme responsible for telomere elongation in cells. It is a basic nuclear protein reverse transcriptase that can add tel...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/25G01N21/64
Inventor 叶汝章张娟全利滕凌叶亚东朱文华潘鹂胡娟郝小江朱兆云路易斯伊格纳罗
Owner 云南路易斯中药现代化工程技术研究中心
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