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Kit for separating genome DNA by using magnetic balls and application thereof

A magnetic bead separation and genome technology, applied in the biological field, can solve the problems of high price and achieve the effects of easy transportation, good safety and high yield

Inactive Publication Date: 2010-08-04
上海鼎国生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Well-known foreign kit manufacturers include Sigma, Promega, Invitrogen, TaKaRa, Whatman, etc. They have developed a series of reagents for materials from different sources such as microorganisms, animal body fluids, blood and tissues, plants, processed products, and genetically modified products. These kits are designed with different extraction methods for different material sources. The operation is simple and efficient, and the DNA quality is high, but the price is expensive

Method used

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  • Kit for separating genome DNA by using magnetic balls and application thereof
  • Kit for separating genome DNA by using magnetic balls and application thereof
  • Kit for separating genome DNA by using magnetic balls and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Preparation of Genomic DNA Separation Kit with Magnetic Beads

[0056] This embodiment includes the following steps:

[0057] 1) Preparation of magnetic balls: uniform, superparamagnetic, single-divergent polymagnetic balls were synthesized using ferric ammonium sulfate and ferrous ammonium sulfate as materials. 9.6g ferric ammonium sulfate, 3.0g ferrous ammonium sulfate were dissolved in 25ml water and 1ml concentrated hydrochloric acid respectively. Put the above solution into a three-necked flask, blow nitrogen gas, stir at 1000rpm, and add 26ml of concentrated ammonia water dropwise for about 30 minutes, continue nitrogen flow, stir for about one hour and stop stirring. After the reaction is complete, wash with water several times to remove tiny particles. During this process, the temperature of the three-necked flask was kept at about 50 degrees. 2) Modified ligand of magnetic spheres: Take a certain amount of magnetic fluid in a three-necked flask. W...

Embodiment 2

[0059] Example 2: Application of this kit to isolate poplar leaf genomic DNA and DNA enzyme digestion detection

[0060] Genomic DNA of poplar leaves was isolated using the kit described in Example 1 and detected by enzyme digestion experiments. This example includes the following steps:

[0061] 1) Genomic DNA extraction: ① Take 50 mg of plant tissue and grind it with liquid nitrogen. ② Add 450 μl of cell lysate and bathe in water at 65°C for 30 minutes. ③Add 300μl binding solution, mix gently, and centrifuge at 12000rpm to remove the precipitate. ④ Add 50 μl of magnetic beads, mix gently, and keep the magnetic beads in suspension for 5 minutes. ⑤ Place the centrifuge tube on the magnetic rack and remove the liquid. ⑥ Add 800 μl of washing solution A, mix gently, put the centrifuge tube on the magnetic rack, and remove the liquid. ⑦Add 800μl washing solution A, mix gently, put the centrifuge tube on the magnetic rack, and remove the liquid. ⑧ Dry at room temperature for...

Embodiment 3

[0072] Example 3 Using this kit to isolate mouse muscle tissue DNA and perform fluorescence quantitative PCR detection

[0073] The kit described in Example 1 was used to isolate mouse muscle tissue genomic DNA and perform fluorescent quantitative PCR experiment detection. This example includes the following steps:

[0074] 1) Genomic DNA extraction: ① Take 50 mg of mouse muscle tissue and grind it with liquid nitrogen. ② Add 450 μl of cell lysate, 10 μl of proteinase K (20 mg / ml), 10 μl of RNase A, and bathe in water at 58°C for 30 minutes. ③Add 300μl binding solution, mix gently, and centrifuge at 12000rpm to remove the precipitate. ④ Add 50 μl of magnetic beads, mix gently, and keep the magnetic beads in suspension for 5 minutes. ⑤ Place the centrifuge tube on the magnetic rack and remove the liquid. ⑥ Add 800 μl of washing solution A, mix gently, put the centrifuge tube on the magnetic rack, and remove the liquid. ⑦Add 800μl washing solution A, mix gently, put the cen...

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Abstract

The invention provides a kit for separating genome DNA by using magnetic balls and application thereof. The kit comprises magnetic balls, a magnetic frame, a genome DNA extraction reagent (lysing solution, binding solution, rinsing solution A, rinsing solution B and eluent) and specifications; the main steps of extracting the gene DNA comprise cell lysis, nucleic acid absorption, impurity removing and nucleic acid elution. The application of the kit for extracting the genome DNA does not need to use large-toxicity organic solvents of phenol, chloroform and the like, has good safety, and simple, fast and time-saving operation, and simultaneously can extract a plurality of samples. The kit can extract the genome DNA from materials of animals, plants, bacteria, fungus, blood, virus, animal source feed stuff, samples in the forensic medicine and the like, the DNA has high yield and purity, and the obtained genome DNA can be used for experiments such as PCR amplification, gene cloning, construction of genomic library, sequence measurement, molecular hybridization, molecular marking and the like. The kit can be stored at the temperature of 4 DEG C, also can be placed at room temperature, and is convenient to transport.

Description

technical field [0001] The invention relates to a kit for extracting genomic DNA from a sample by using magnetic beads, which belongs to the field of biotechnology and relates to the composition and application of the kit. Background technique [0002] Life is determined by the genome. The vast majority of genomes, including those of all cellular life forms, are composed of DNA. Therefore, in order to carry out sequencing, hybridization and gene expression, obtaining high molecular weight and high purity DNA is a very important prerequisite, and DNA extraction has become one of the most important and basic operations in molecular biology experimental techniques. [0003] In eukaryotes, 95% of genomic DNA exists in the nucleus, and 5% is distributed in organelles such as mitochondria and chloroplasts. The vast majority of plant tissue is nuclear DNA, which is combined with histones and non-histones and exists in the nucleus in the form of nucleoproteins (ie, chromatin or ch...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68
Inventor 周卫东
Owner 上海鼎国生物技术有限公司
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