Kit for recovering DNA (Deoxyribose Nucleic Acid) by using column method agarose gel and method for purifying DNA

A technology of agarose gel and kit, which is applied in the field of kits for recovery of DNA from column-type agarose gel, which can solve the problems of low recovery, time-consuming and economic cost, and impact on recovery efficiency, and achieve high recovery efficiency , Easy to operate, wide range of practical effects

Pending Publication Date: 2022-03-15
艾柏森(江苏)生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the recovery of DNA by the column method also has disadvantages, such as low recovery. If a large amount of nucleic acid needs to be recovered, more columns are required, which consumes time and economic costs and affects the recovery efficiency.

Method used

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  • Kit for recovering DNA (Deoxyribose Nucleic Acid) by using column method agarose gel and method for purifying DNA
  • Kit for recovering DNA (Deoxyribose Nucleic Acid) by using column method agarose gel and method for purifying DNA
  • Kit for recovering DNA (Deoxyribose Nucleic Acid) by using column method agarose gel and method for purifying DNA

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Embodiment 1

[0024] The kit for recovering DNA from agarose gel by column method disclosed in this example includes a sol solution, a DNA adsorption column and a washing solution. Containing concentration is 4.5mol / L guanidine isothiocyanate, 0.2mol / L acetic acid-potassium acetate buffer solution and 0.03g / L phenol red in the sol solution, the pH value of the acetic acid-potassium acetate buffer solution used to configure the sol solution is 4.8; The DNA adsorption column adopts a 3-layer silica gel membrane column; the rinsing solution contains 0.3mol / L NaCl, 0.003mol / L EDTA, 0.03mol / L Tris-HCl buffer and 75% absolute ethanol.

[0025] When using the kit of this embodiment to purify and recover DNA, the specific operations are as follows:

[0026] A. Select 150bp, 750bp and 5000bp DNA samples for agarose gel electrophoresis. After the end, use a clean scalpel to cut out the target DNA band under ultraviolet light and transfer it to a 2mL centrifuge tube.

[0027] B. The cut gel piece is ...

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Abstract

The invention provides a kit for recovering DNA (Deoxyribose Nucleic Acid) by using agarose gel through a column method. The kit comprises a sol solution, a DNA adsorption column and a rinsing solution, the sol solution consists of guanidine isothiocyanate with the concentration of 4.1 mol/L to 4.8 mol/L, an acetic acid-potassium acetate buffer solution with the concentration of 0.1 mol/L to 0.5 mol/L and phenol red with the concentration of 0.01 g/L to 0.05 g/L; a DNA adsorption column adopts a three-layer silica gel membrane column; the rinsing liquid is prepared from 0.1 mol/L to 0.5 mol/L of NaCl, 0.001 mol/L to 0.005 mol/L of EDTA (Ethylene Diamine Tetraacetic Acid), 0.01 mol/L to 0.05 mol/L of a Tris-HCl buffer solution and 75 percent of absolute ethyl alcohol. The invention also provides a method for purifying and recovering DNA by using the kit for recovering DNA by using the column method agarose gel. The agarose gel DNA recovery kit and the method for purifying and recovering DNA by using the kit have the advantages of low cost, stability and high efficiency.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for recovering DNA from a column-type agarose gel and a method for purifying DNA. Background technique [0002] Agarose gel recovery of DNA is one of the most common techniques in gene cloning experiments. Existing nucleic acid purification methods include low-melting point agarose gel recovery, DEAE cellulose membrane disc method, dialysis bag electroelution method, glass milk method, agarose enzymatic method, kit column recovery, magnetic bead method, etc., but Some methods have disadvantages such as long recovery time, cumbersome operation, and high cost of reagents. Column recovery of DNA is still the most widely used method with high operating efficiency and low recovery cost. [0003] Currently, the most commonly used agarose gel recovery method in the laboratory is the column method. However, the recovery of DNA by the column method also has disadvantages, such as low...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/10
Inventor 谢朋辉贺建望余晓锐
Owner 艾柏森(江苏)生物科技有限公司
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