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A method of inhibiting dna enzyme cleavage

A DNase and double-enzyme cleavage technology, applied in the field of molecular biology, can solve the problems of poor temperature control and inability to completely inhibit enzyme cleavage, and achieve the effects of convenient operation, easy acquisition and reasonable cost.

Active Publication Date: 2021-10-12
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some enzymes also have enzymatic activity at high temperature, the temperature is not easy to control, and some cannot completely inhibit enzyme digestion

Method used

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  • A method of inhibiting dna enzyme cleavage
  • A method of inhibiting dna enzyme cleavage
  • A method of inhibiting dna enzyme cleavage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] (1) Preparation of monolayer or multilayer MoS 2 The specific steps are as follows:

[0028] This example prepares 2mg / mL of MoS 2 , Weigh 10 mg of the initial molybdenum disulfide crystal, dissolve it in a glass bottle filled with 5mL ultrapure water, and ultrasonicate for ≥2h, wherein the original molybdenum disulfide crystal is obtained by mechanical exfoliation, that is, the material with a layered structure and Other harder substances are subjected to physical surface friction to obtain a thin layer or monolayer material, and the average diameter of the nanomaterial is maintained at 176nm.

[0029] figure 1 Schematic diagram of the molecular structure of molybdenum disulfide. The monolayer structure of the material consists of a quasi-2D network of covalently bonded S-Mo-S hexahedrons.

[0030] (2) Preparation of Escherichia coli containing the target plasmid:

[0031] a. Preparation of competent cells: Take out the frozen Escherichia coli DH5α strain from th...

Embodiment 2

[0046] (1) This example prepares 1 mg / mL MoS 2 , Weigh 10mg of the initial molybdenum disulfide crystal, dissolve it in a glass bottle filled with 10mL ultrapure water, and ultrasonicate for 3h, wherein the original molybdenum disulfide crystal is obtained by mechanical exfoliation, that is, the material with a layered structure and other Harder substances are subjected to physical surface friction to obtain thin or monolayer materials, and the average diameter of the nanomaterials is maintained at 214nm.

[0047] (2) Preparation of Escherichia coli containing the target plasmid:

[0048] a. Preparation of competent cells: the steps are the same as in a in Example 1 (2).

[0049] b. Transformation: take the competent cells out of the -70°C refrigerator, thaw on ice, and work in ultra-clean

[0050] In Taichung, take 1 μL of PB2GW7.0 plasmid and add 9 μL of ultrapure water to dilute it, then add it to 100 μL of competent cells, place on ice for 30 minutes, heat shock (water b...

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Abstract

The invention provides a method for inhibiting DNA enzymatic cleavage, which belongs to the field of molecular biology; the present invention controls the technical parameters of the reaction by adding molybdenum disulfide nanomaterials into the enzymatic cleavage reaction system to achieve the purpose of inhibiting DNA enzymatic cleavage. Using enzyme cleavage experiments and cross-combination of nanomaterials, inhibition of enzyme cleavage is expected to be applied to cell signaling pathways, preventing the activity of related enzymes, resulting in certain signals not being conveyed smoothly, providing a new research direction and related theoretical reference for biomedicine , In addition, MoS in the present invention 2 Nanomaterials are stable in properties, easy to prepare, high in yield, resistant to acid and alkali, low in preparation cost, and have certain application and development prospects.

Description

technical field [0001] The invention provides a method for inhibiting DNA enzyme cutting, which belongs to the field of molecular biology. Background technique [0002] Enzyme cleavage technology generally refers to directional enzyme cleavage technology, and directional enzyme cleavage technology specifically refers to the use of restriction endonucleases to cut DNA fragments. Due to the specificity of enzymes, that is, an enzyme can only recognize a specific deoxynuclear Nucleotide sequence, so this specific enzyme can be used to cut the corresponding DNA fragment, and then achieve the purpose of directional cutting. Directed enzyme digestion technology generally uses single enzyme digestion and double enzyme digestion methods, especially double enzyme digestion is a commonly used method in biological experiments. Enzyme digestion experiments exist in various fields such as biology, chemistry, and medicine, and are indispensable experimental links. In the process of enzy...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/22
CPCC12N9/22
Inventor 高力夏妮唐琦李国辉吴鹏
Owner JIANGSU UNIV
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