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Method for inhibiting DNA digestion

A DNase and double-enzyme digestion technology, applied in the field of molecular biology, can solve the problems of incomplete inhibition of enzyme digestion, poor temperature control, etc., and achieve the effects of easy acquisition, convenient operation, and simple principle

Active Publication Date: 2019-01-11
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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  • Method for inhibiting DNA digestion
  • Method for inhibiting DNA digestion
  • Method for inhibiting DNA digestion

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 1mg / 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 ba...

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Abstract

The invention provides a method for inhibiting DNA digestion, and belongs to the field of molecular biology. According to the method, a molybdenum disulfide nano-material into a digestion reaction system to control the technical parameter of the reaction, and the aim of inhibiting DNA digestion can be achieved; and cross-linking of the digestion experiment and the nano-material is utilized, digestion inhibition is expected to be applied to a cell signal pathway, the activity of related enzyme can be stopped, certain signals cannot be delivered smoothly, and a new research direction and relatedtheoretical reference foundation are provided to biomedicine. Furthermore, the MoS2 nano-material has stable property, is easy to prepare, has high yield, acid and alkali prevention and relatively low preparation cost, and has a certain application development prospect.

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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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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