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Cell lysis solution and application thereof

A technology of cell lysate and alkaline solution, applied in the field of cell lysate, can solve the problem that a sufficient amount of DNA cannot be extracted from the cell lysate, and achieve the effects of saving nucleic acid extraction time, simple operation steps and low cost.

Pending Publication Date: 2021-04-09
广州源井生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to overcome the deficiency that the existing cell lysate cannot extract a sufficient amount of DNA from a small amount of cells for PCR genotype identification, the purpose of the first aspect of the present invention is to provide a cell lysate

Method used

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  • Cell lysis solution and application thereof
  • Cell lysis solution and application thereof
  • Cell lysis solution and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 A kind of cell lysate

[0039] A cell lysate, composed of A component and B component, the raw materials of A component and B component are composed as follows:

[0040] Component A: 5mmol / L NaOH, 0.2mmol / L sodium lauryl sulfate;

[0041] Component B: 5mmol / L Tris, 10mmol / L KCl, 1mmol / L EDTA, pH7.0;

[0042] When used, the volume ratio of component A to component B is 3:1.

Embodiment 2

[0043] Embodiment 2 A kind of cell lysate

[0044] A cell lysate, composed of A component and B component, the raw materials of A component and B component are composed as follows:

[0045] Component A: 100mmol / L NaOH, 2.5mmol / L sodium lauryl sulfate;

[0046] Component B: 200mmol / L Trisl, 150mmol / L KCl, 20mmol / L EDTA, pH8.0;

[0047] When used, the volume ratio of component A to component B is 3:1.

Embodiment 3

[0048] A kind of cell lysate of embodiment 3

[0049] A cell lysate, composed of A component and B component, the raw materials of A component and B component are composed as follows:

[0050] Component A: 200mmol / L NaOH, 3.5mmol / L sodium lauryl sulfate;

[0051] Component B: 500mmol / L Tris, 1000mmol / L KCl, 100mmol / L EDTA, pH9.0;

[0052] When used, the volume ratio of component A to component B is 3:1.

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PUM

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Abstract

The invention discloses a cell lysis solution and application thereof. The cell lysis solution is composed of a component A and a component B, wherein the component A is composed of 3-600mmol / L of an alkali solution and 0.1-7mmol / L of SDS; the component B is composed of 3-600mmol / L of Tris, 8-1100mmol / L of chlorate and 0.5-110mmol / L of EDTA, and the pH value is 7-10; and the volume ratio of the component A to the component B is (3-50): 1 during use. The cell lysis solution does not contain chloroform or other toxic reagents, is composed of common reagents, is low in cost and simple in operation steps, can quickly release nucleic acid in cells, and greatly saves the nucleic acid extraction time; and the cell lysis solution can extract sufficient nucleic acid from trace cells (the number of cells is less than or equal to 2 * 10< 4 >) for PCR identification.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a cell lysate and its application. Background technique [0002] Gene editing technology is a technology that precisely manipulates the DNA sequence in cells to change the fate of cells and the characteristics of organisms. This technology provides an important tool for improving human understanding of genetics and the treatment of genetic diseases. In particular, the invention of CRISPR-Cas9 gene editing technology has brought revolutionary changes to the development of gene editing technology. CRISPR-Cas9 technology has made gene editing operations simple and fast, but it is still time-consuming and laborious to screen gene-edited mutants, especially cell gene-edited mutants. In order to obtain a homozygous gene-edited cell line, it is necessary to perform monoclonal screening of the gene-edited cells, and inoculate a single cell into a 96-well microplate for culture. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/686
CPCC12Q1/6806C12Q1/686C12Q2527/125C12Q2565/125
Inventor 郑丹丹黄文静叶海峰
Owner 广州源井生物科技有限公司