Improved apoptotic DNA ladder extraction kit

A kit and apoptosis technology, applied in the field of medical detection reagents, can solve the problems of cumbersome steps, long lysis time, irregular length, etc., and achieve the effects of avoiding environmental pollution, simple operation process and reducing experimental cost.

Inactive Publication Date: 2014-01-01
张根
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The advantage of this method is that the method of operation is simple and easy to master, but the disadvantage is that the extracted product is mixed with DNA of irregular length from the total genome and necrotic cells in addition to apoptotic DNA.
The effect of DNA gel electrophoresis run out in this way can be imagined, the discrimination between apoptotic DNA ladders (between bands) is very poor, which ultimately leads to poor experimental results
(2) Or although it is not separated and eluted by a centrifugal column, becau

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Improved apoptotic DNA ladder extraction kit
  • Improved apoptotic DNA ladder extraction kit
  • Improved apoptotic DNA ladder extraction kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 An improved apoptotic DNA ladder extraction kit, including

[0051] - cell lysate, used to extract the apoptotic nucleosomes in the apoptotic cells within 10 seconds;

[0052] The cell lysate consists of 0.121g~0.200g Tris, 2.340~3.000g NaCl, 0.029g~0.040gEDTA, 1mL~2mL Tritan-x.

[0053] —Extract reagent enzyme A solution and B solution, which are used to remove RNase and nucleosomal protein mixed in apoptotic nucleosomes;

[0054] Solution A consists of 250 μL of an aqueous solution containing 250 μg of RNase;

[0055] Solution B consisted of an aqueous solution containing 500 μg proteinase K in 250 μL.

[0056] - ammonium acetate solution;

[0057] The ammonium acetate solution consisted of an aqueous solution containing 790 mg ammonium acetate in 1 mL.

[0058] - DNA ladder solution;

[0059] The DNA ladder solution consists of 1.667mL of TE buffer and 0.333mL of an aqueous solution containing 4.4g of EDTA, 250mg of bromophenol blue, and 250mg of...

Embodiment 2

[0069] Example 2 An improved apoptotic DNA ladder extraction kit, including

[0070] - cell lysate, used to extract the apoptotic nucleosomes in the apoptotic cells within 10 seconds;

[0071] The cell lysate consists of 0.121g~0.200g Tris, 2.340~3.000g NaCl, 0.029g~0.040gEDTA, 1mL~2mL Tritan-x.

[0072] —Extract reagent enzyme A solution and B solution, which are used to remove RNase and nucleosomal protein mixed in apoptotic nucleosomes;

[0073] Solution A consists of 250 μL of an aqueous solution containing 250 μg of RNase;

[0074] Solution B consisted of an aqueous solution containing 500 μg proteinase K in 250 μL.

[0075] - ammonium acetate solution;

[0076] The ammonium acetate solution consisted of an aqueous solution containing 790 mg ammonium acetate in 1 mL.

[0077] - DNA ladder solution;

[0078] The DNA ladder solution consists of 1.667mL of TE buffer and 0.333mL of an aqueous solution containing 4.4g of EDTA, 250mg of bromophenol blue, and 250mg of ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to an improved apoptotic DNA ladder extraction kit, which comprises a cell lysis buffer used for extracting an apoptotic nucleosome of an apoptotic cell within 10 seconds, an extraction reagent enzyme A solution and an extraction reagent enzyme B solution both used for removing an RNA enzyme and nucleosome protein mixed in the apoptotic nucleosome, an ammonium acetate solution formed by an aqueous solution in which 1 mL aqueous solution contains 790 mg ammonium acetate, and a DNA ladder solution formed by a TE buffer solution of 1.667 mL and an aqueous solution in which 0.333 mL aqueous solution contains 4.4 g EDTA, 250 mg bromophenol blue and 250 mg Xylene Cyanol FF. The improved apoptotic DNA ladder extraction kit used for detecting ligand has the characteristics of quickness, high sensitivity (cells with the apoptosis rate more than 10% can be detected), capability of qualitative detection, and strong specificity.

Description

technical field [0001] The invention relates to the technical field of medical detection reagents, in particular to an improved apoptotic DNA ladder extraction kit. Background technique [0002] Apoptosis refers to the autonomous and orderly death of cells controlled by genes in order to maintain a stable internal environment. It plays an important role in the evolution of organisms, the stability of the internal environment, and the development of multiple systems. The main features of apoptosis are cell shrinkage, changes in cell membrane structure, dense nuclear chromatin, and endogenous nuclease cutting of genomic DNA. [0003] Early cell apoptosis detection is morphological observation method: (1) HE staining and light microscope observation: apoptotic cells are round, with deeply stained nuclei, concentrated cytoplasm, clumps of chromatin, and "budding" phenomenon on the cell surface. ⑵ Acridine orange (AO) staining, observation by fluorescence microscope: living cel...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/10
Inventor 张根赵晓辉曾新刘宾袁毅陈明月
Owner 张根
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap