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Method for recovery of DNA by using magnetic nanocomposite and reagent kit therefor

A composite material and magnetic nanotechnology, applied in the biological field, achieves the effect of simple method, good biocompatibility and prolonging service life

Inactive Publication Date: 2007-05-30
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Currently, there is no technology for recovering DNA using magnetic nanocomposites

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Preparation of Magnetic Nanocomposites

[0046] Weigh 54.90g of FeCl 3 ·6H 2 O and 20.20 g FeCl 2 4H 2 O was dissolved in 200ml of water through nitrogen and deoxygenation to obtain a mixed solution. Take 800ml of water in a 1L round-bottomed flask, pass nitrogen to remove oxygen, add 50ml of concentrated ammonia water with a concentration of 25-28% by mass, and quickly pour the above-mentioned iron salt mixed solution into it under vigorous stirring. Reaction 1h. After the reaction is completed, use a 0.1T permanent magnet to separate the black solid from the reaction solution, and wash the obtained solid with high-purity water for 3 to 5 times to obtain Fe with a particle size of 6 to 10 nm. 3 o 4 Nanoparticles.

[0047] Weigh 5.0g Na 2 SiO 3 (SiO 2 Content is 45%) be dissolved in 1000ml water, be 9.6 with the pH adjustment solution of cationic resin, add 2g above-mentioned synthetic Fe wherein 3 o 4 Nanoparticles. At room temperature, the reaction was ca...

Embodiment 2

[0050] DNA Recovery Kit

[0051] Preparation of DNA recovery kit using magnetic composite material, which includes:

[0052] 0.5ml concentration is the magnetic nanocomposite material solution prepared in the embodiment 1 of 20mg / ml

[0053] 100ml solution, its composition is 6mol / LNaI solution, 0.5ml Tris-HCl, pH=6.4, 25mmol / l EDTA;

[0054] 100ml washing liquid II, its component is 1.5mol / l sodium acetate;

[0055] 10ml eluent, deionized water.

Embodiment 3

[0057] DNA Recovery Kit

[0058] Preparation utilizes the DNA recovery kit of magnetic composite material, and the magnetic nanocomposite material solution in it, washing liquid II are the same as embodiment 2, also include:

[0059] 100ml solution, its components are 4mol / L NaCl solution, 0.5ml Tris-HCl, pH=6.4, 45mmol / l EDTA, 2.5% Triton;

[0060] 100ml washing liquid I, its composition is the NaClO of 4mol / l 4 Solution, 0.5mol / l Tris-HCl, pH=6.7, 25mmol / l EDTA;

[0061] 10ml eluent, its composition is 10mmol / l Tris-HCl.

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Abstract

The invention discloses a recycling method of DNA and agent box through magnetic nanometer composite material in the biological technical domain, which comprises the following steps: (a) blending magnetic nanometer composite material and recycled sample; forming solid-liquid dispersing suspension; (b) adsorbing DNA to separate magnetic nanometer composite material from solid phase; obtaining magnetic nanometer composite material with adsorbed DNA; (c) washing impurity; evolving DNA from magnetic nanometer composite material. The magnetic nanometer composite agent box to recycle DNA possesses magnetic nanometer composite material solution, dissolving liquid, washing liquid I and washing liquid II and eluent.

Description

technical field [0001] The present invention relates to the field of biotechnology. More specifically, the present invention relates to a method for recovering DNA using magnetic nanoparticles and a kit for recovering DNA. Background technique [0002] The recovery of DNA has a wide range of applications, such as recovery of high-purity DNA from DNA polymerase chain reaction (PCR) amplification products, polyacrylamide gel, agarose gel, digested biological samples for genetic fingerprint identification, gene Sequencing, library construction, PCR testing, etc. DNA recovery technology is a key technology in molecular biology experiments. The efficiency and purity of recovered DNA will directly affect the success of subsequent experiments. Therefore, developing fast, efficient, simple and low-cost DNA recovery methods has always One of the goals of academics. [0003] At present, the main DNA recovery methods are chemical method, glass bead method and column separation metho...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 杨百全王刚杨文胜
Owner JILIN UNIV
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