Method of shearing DNA molecules and application thereof

A technology of DNA molecules and nanoparticles, which is applied in the field of shearing DNA molecules, can solve the problems of high temperature requirements, inability to obtain uniform structures, and non-specificity of target sequences, and achieve high specificity and biocompatibility Effect

Active Publication Date: 2018-12-14
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the commonly used methods for shearing DNA molecules are as follows: DNA restriction enzyme digestion, sonication, hydrodynamic shearing, etc. Enzyme cleavage still requires specific biological enzymes, which has the disadvantage of high pH and temperature requirements; sonication and hydrodynamic shearing methods have the disadvantage of no specificity to the target sequence
[0005] Therefore, none of the above-mentioned methods can obtain DNA fragments with uniform structure and high specificity in sequence and position. We urgently need to find a DNA fragment that can obtain uniform structure and high specificity in sequence and position. methods to meet the needs of genetic research

Method used

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  • Method of shearing DNA molecules and application thereof
  • Method of shearing DNA molecules and application thereof
  • Method of shearing DNA molecules and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: the synthesis of precursor

[0046] Under the protection of nitrogen, 4 mL of 0.5 M H 2 SO 4 Added to 0.05g Al 2 Te 3 solution, the precursor gas is obtained.

Embodiment 2

[0047] Example 2: Synthesis of Chiral Semiconductor Nanoparticles

[0048] 0.985g of Cd(ClO 4 ) 2 ·6H 2 After the D-type cysteine ​​of 0 and 3mL 1M is added in 125mL water, pass into the precursor gas that embodiment 1 obtains, adopt NaOH to regulate pH to be 12, the precursor gas adopts nitrogen as carrier gas, and flow velocity control is 100mL / min, The solution was heated to 110° C. while stirring, and kept for 8 h, and then, isopropanol was added in a volume ratio of 1:1 to the obtained nanoparticle solution, and then the mixture was centrifuged at 10,000 rpm for 5 min. The pellet was then resuspended in PBS buffer (0.01M, pH 7.4) to obtain chiral semiconductor nanoparticles. ( figure 1 TEM image of chiral semiconductor nanoparticles, figure 2 is the circular dichroism spectrum of chiral semiconductor nanoparticles)

Embodiment 3

[0049] Example 3: Synthesis of Chiral Semiconductor Nanoparticles

[0050] 0.985g of Cd(ClO 4 ) 2 ·6H 2After the L-type cysteine ​​of 0 and 3mL 1M is added in 125mL water, pass into the precursor gas that embodiment 1 obtains, adopt NaOH to regulate pH to be 12, the precursor gas adopts nitrogen as carrier gas, and flow velocity control is 100mL / min, The solution was heated to 110° C. while stirring, and kept for 8 h, and then, isopropanol was added in a volume ratio of 1:1 to the obtained nanoparticle solution, and then the mixture was centrifuged at 10,000 rpm for 5 min. The pellet was then resuspended in PBS buffer (0.01M, pH 7.4) to obtain chiral semiconductor nanoparticles. ( figure 1 TEM image of chiral semiconductor nanoparticles, figure 2 is the circular dichroism spectrum of chiral semiconductor nanoparticles)

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Abstract

The invention discloses a method of shearing DNA molecules and application thereof and belongs to the technical field of gene engineering. The method comprises the following steps: firstly, mixing chiral semiconductor nanoparticles with DNA molecules for incubation; and then irradiating the incubated mixture of the chiral semiconductor nanoparticles and the DNA molecules under circularly polarizedlight to obtain sheared DNA molecules. A lot of DNA fragments which are uniform in structure and high in specificity in sequence and site can be acquired quickly and have huge application potential in the field of gene engineering.

Description

technical field [0001] The invention relates to a method for cutting DNA molecules and an application thereof, belonging to the technical field of genetic engineering. Background technique [0002] Obtaining DNA fragments is a key step for technologies such as gene sequencing, gene analysis, and gene editing. For gene sequencing, it can effectively obtain target fragments; for gene analysis, it can obtain single gene information; for gene editing, it can selectively extract or copy gene sequences. Therefore, the shearing of DNA molecules is an indispensable pretreatment step in the field of genetic engineering. [0003] With the continuous deepening of research in the field of genes, the demand for obtaining DNA fragments with uniform structure and high specificity in sequence and position is becoming greater and greater. [0004] At present, the commonly used methods for shearing DNA molecules are as follows: DNA restriction enzyme digestion, ultrasonic degradation, hydro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/10
Inventor 匡华孙茂忠胥传来徐丽广马伟刘丽强宋珊珊吴晓玲
Owner JIANGNAN UNIV
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