Improved apoptotic DNA ladder extraction kit

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...

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, because the lysate uses a genome lysate with strong lysis, and the lysis time is long, it is bound to contain a large amount of genomic DNA in the extracted product, which will be to a certain extent Under masking the effect of the extracted apoptotic DNA ladder

The disadvantages of this are that firstly, it increases the cost of the test, and secondly, the steps are cumbersome and highly toxic, and it is generally operated in a fume hood

Moreover, due to the process of protein extraction, the apoptotic DNA ladder has a certain loss

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