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Method for purifying DNA by using gold magnetism particles

A gold magnetic particle and magnetic separation technology, which is applied in DNA preparation, recombinant DNA technology, etc., can solve the problems of low DNA purification efficiency, long operation time, and poor purity, and achieve large fixed capacity, long drying time, and high purification rate Effect

Active Publication Date: 2008-04-23
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a method for purifying DNA with gold magnetic particles, which solves the shortcomings of the prior art such as low DNA purification efficiency, poor purity, and long operating time

Method used

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  • Method for purifying DNA by using gold magnetism particles
  • Method for purifying DNA by using gold magnetism particles
  • Method for purifying DNA by using gold magnetism particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 is the specific process of rapid purification of genomic DNA from whole blood:

[0030] 1] Pretreatment of gold magnetic particles:

[0031] 1.1] Fully mix the gold magnetic particles to ensure that the magnetic particles are completely suspended in the solution;

[0032] 1.2] Take 100 μl of gold magnetic particles in a 1.5ml centrifuge tube, place the centrifuge tube on a magnetic separator for magnetic separation until the supernatant is clear; the separation can be completed within 1 min, and the supernatant is removed;

[0033] 1.3] Separate the centrifuge tube from the magnetic separator. Add 200 μl of pretreatment buffer to the tube, mix with a pipette, then put it back on the magnetic separator, and remove the supernatant;

[0034] 1.4] Repeat steps 1.3] to 1.4] twice. Then add 200 μl coupling buffer to the centrifuge tube and mix thoroughly.

[0035] 2] DNA purification steps:

[0036] 2.1] Take 200 μl whole blood and add it to a 1.5ml centrifu...

Embodiment 2

[0045] Embodiment 2 is the specific process of purifying plasmid DNA from bacteria:

[0046] 1] Pretreatment of gold magnetic particles:

[0047] 1.1] Fully mix the gold magnetic particles to ensure that the magnetic particles are completely suspended in the solution;

[0048] 1.2] Take 500 μl of gold magnetic particles in a 1.5ml centrifuge tube, place the centrifuge tube on a magnetic separator for magnetic separation until the supernatant is clear; the separation can be completed within 1 min, and the supernatant is removed;

[0049] 1.3] Separate the centrifuge tube from the magnetic separator. Add 200 μl of pretreatment buffer to the tube, mix with a pipette, then put it back on the magnetic separator, and remove the supernatant;

[0050] 1.4] Repeat steps 1.3] to 1.4] twice; then add 200 μl coupling buffer to the centrifuge tube and mix well;

[0051] 2] DNA purification steps:

[0052] 2.1] Take 1ml of the overnight culture solution, centrifuge at the highest speed ...

Embodiment 3

[0060] Embodiment 3 is the specific process of purifying genomic DNA from tissue:

[0061] 1.1] Take 10mg~100mg tissue high-speed homogenate and add it to a 1.5ml centrifuge tube;

[0062] 1.2] Add 30 μl ~ 300 μl lysis buffer I and 60 μl ~ 600 μl lysis buffer II to the tube, then add 1 μl ~ 10 μl RNase A solution, blow and mix with a pipette; stand at room temperature for 5 minutes;

[0063] 1.3] Add 45 μl ~ 450 μl lysis buffer III to the tube, and mix with a pipette; ice bath for 10 minutes;

[0064] 1.4] Add the pretreated gold magnetic particles into the tube, gently blow and mix with a pipette; let it stand at room temperature for 3 minutes; place the centrifuge tube on a magnetic separator for magnetic separation for 5 minutes, and remove the supernatant;

[0065] 1.5] Remove the centrifuge tube from the magnetic separator; add 200 μl ~ 1000 μl of washing buffer to the tube and mix with a pipette. Place the centrifuge tube back on the magnetic separator for magnetic sep...

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Abstract

The DNA purifying magnetic metal particle process includes the following steps: 1. preparing liquid sample containing DNA; 2. mixing magnetic metal particle, which is composite magnetic particle comprising a core of magnetic nanometer particle and a shell of Au, Ag or other noble metal, and has average size of 0.05-100 microns, and the liquid sample to form magnetic metal particle-DNA composition; 3. applying external magnetic field to separating DNA adsorbing magnetic metal particle from other components in the liquid sample; and 4. eluting DNA adsorbed onto the surface of the magnetic material. The process can separate and purify DNA effectively from various kinds of sample, and has the advantages of high purification rate, high DNA integrity, high purity and convenient operation.

Description

technical field [0001] The invention relates to a method for purifying DNA. Background technique [0002] DNA purification is a key technology in molecular biology experiments and the premise and basis of downstream experiments. The quality of purification will directly affect the success of future experiments, such as PCR amplification, enzyme digestion reaction, blood, saliva, etc. Purified DNA from samples is used for paternity testing, genetic identification of criminal offenses, and purification of DNA from various tissues to construct libraries, etc. Therefore, seeking fast, efficient, simple and low-cost DNA purification methods has always been the direction of people's attention. [0003] Several DNA extraction methods commonly used at present have some shortcomings. Although the phenol-chloroform purification method is suitable for most biological samples, the experimental operation time is long, the steps are cumbersome, and it is easy to cause DNA contamination....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 崔亚丽陈超曹树辉
Owner XIAN GOLDMAG NANOBIOTECH
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