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Nucleic acid extraction and purification method based on nanometer magnetic beads and kit

A nano-magnetic bead and purification method technology, which is applied in DNA preparation, recombinant DNA technology, etc., can solve the problems of long extraction time, complicated extraction steps, and low sample purity, and achieve improved extraction throughput, low cost, and reduced errors The effect of chance

Active Publication Date: 2014-05-28
苏州天隆生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The existing magnetic bead extraction method needs to rely on proteinase K, and the extraction steps are more complicated, so the required extraction time will be relatively long, and the extracted sample also has the problem of not too high purity, so the purified sample Can not fully meet the follow-up experimental requirements

Method used

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  • Nucleic acid extraction and purification method based on nanometer magnetic beads and kit
  • Nucleic acid extraction and purification method based on nanometer magnetic beads and kit
  • Nucleic acid extraction and purification method based on nanometer magnetic beads and kit

Examples

Experimental program
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Effect test

Embodiment 1

[0026] 1) Put 200ul of EDTA anticoagulant blood into a centrifuge tube, add 600ul of lysis buffer into the centrifuge tube, mix at room temperature for 3 minutes, then place the centrifuge tube on a magnetic stand, separate by magnetic force for 20 seconds, and discard the supernatant;

[0027] Wherein, the ingredients added in the lysis buffer are: sodium iodide 2M, guanidine hydrochloride 2.5M, EDTA 10mM, Tween-203%, nano magnetic beads 8%, SDS 2%, isopropanol 35%, and the remaining ingredients are water , and the pH of the lysis buffer was adjusted to 7.4.

[0028] 2) Continue to add 400ul of washing buffer solution to the centrifuge tube, mix for 1 minute, place the centrifuge tube on a magnetic stand, magnetically separate for 20 seconds, discard the supernatant, open the cover and let it cool for 1 minute;

[0029] Wherein, the ingredients added to the washing buffer are: EDTA 5mM, Tris-cl 150mM, ethanol 75%, and the remaining ingredients are water, and the pH value of t...

Embodiment 2

[0036] 1) Put 100ul of serum containing hepatitis C virus (HCV virus) into a centrifuge tube, add 300ul of lysis buffer into the centrifuge tube, mix at room temperature for 3 minutes, then place the centrifuge tube on a magnetic stand, magnetically separate for 20s, and absorb Discard the supernatant;

[0037] Wherein, the ingredients added in the lysis buffer are: sodium iodide 2M, guanidine hydrochloride 2.5M, EDTA 10mM, Tween-203%, nano magnetic beads 8%, SDS 2%, isopropanol 35%, and the remaining ingredients are water , and the pH of the lysis buffer was adjusted to 7.4.

[0038]2) Continue to add 400ul of washing buffer solution to the centrifuge tube, mix for 1 minute, place the centrifuge tube on a magnetic stand, magnetically separate for 20 seconds, discard the supernatant, open the cover and let it cool for 1 minute;

[0039] Wherein, the ingredients added to the washing buffer are: EDTA 5mM, Tris-cl 150mM, ethanol 75%, and the remaining ingredients are water, and ...

Embodiment 3

[0050] 1) Take 100ul of serum containing hepatitis B virus (HBV virus) and place it in a centrifuge tube, add 300ul of lysis buffer into the centrifuge tube, mix at room temperature for 3 minutes, then place the centrifuge tube on a magnetic stand, magnetic separation for 20s, and absorb Discard the supernatant;

[0051] 2) Continue to add 400ul of washing buffer solution to the centrifuge tube, mix for 1 minute, place the centrifuge tube on a magnetic stand, magnetically separate for 20 seconds, discard the supernatant, open the cover and let it cool for 1 minute;

[0052] 3) Finally, add 100ul of elution buffer to the centrifuge tube, mix for 1 minute, place the centrifuge tube on a magnetic stand, and separate by magnetic force for 20 seconds, transfer the supernatant to another clean, enzyme-free centrifuge tube, The final extracted and purified HBV viral nucleic acid RNA solution can be obtained by storing at -20°C.

[0053] Wherein, the lysis buffer, washing buffer and ...

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Abstract

The invention discloses a nucleic acid extraction and purification method based on nanometer magnetic beads, comprising the following steps: mixing a biological sample and a lysis buffer to make nanometer magnetic beads in the lysis buffer and nucleic acid DNA / RNA which moves into the lysis buffer form a magnetic bead-nucleic acid compound; transferring the compound under the action of a magnetic field to a washing buffer to wash off impurities on the magnetic bead-nucleic acid compound; and transferring the washed magnetic bead- nucleic acid compound under the action of the magnetic field to an elution buffer so as to elute and recover nucleic acid. The nanometer magnetic beads used in the invention have advantages of uniform size, smooth surface, large surface area ratio, high adsorption capacity of nucleic acid, fast magnetic response speed and rapid separation, and can be stored together with the lysis buffer at room temperature for a long time. The extracted nucleic acid DNA / RNA has high purity, is complete and can be directly used for follow-up detection. The method provided by the invention has shorter nucleic acid extraction time than a general magnetic bead method by the use of a nucleic acid extraction reagent, is more suitable for automation and is adopted to realize high-flux nucleic acid DNA / RNA extraction.

Description

technical field [0001] The invention relates to the field of extracting and purifying nucleic acid by a magnetic bead method, in particular to a method for extracting and purifying nucleic acid based on nano magnetic beads and a kit. Background technique [0002] Nucleic acid includes two types of deoxyribonucleic acid (RNA) and ribonucleic acid (DNA). It is known that nucleic acid is the genetic material of all organisms. It mainly exists in the nucleus in cells and in the viral capsid in viruses. At present, the extraction and purification of nucleic acids are inseparable in all fields of biomedicine. The application of biological nanomaterials, rapid and high-throughput separation and purification to obtain high-purity and complete target nucleic acid DNA / RNA is the basis of biology. [0003] Because of this, there are various nucleic acid extraction methods, such as phenol / chloroform and other organic solvent extraction methods, chelating resin method, glass powder adsor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 李明李红东苗保刚彭年才倪晓龙李政
Owner 苏州天隆生物科技有限公司
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