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Method for purifying total nucleic acid by using gold-magnetic particles

A gold magnetic particle, nucleic acid technology, applied in DNA preparation, recombinant DNA technology and other directions, can solve the problems of inability to purify total nucleic acid at the same time, complex and other problems, achieve the effect of simple operation, fast purification process, and avoid cross-contamination

Active Publication Date: 2010-06-09
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This process is not only complicated, but also belongs to specific separation, and the total nucleic acid of biological samples cannot be purified at the same time

Method used

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  • Method for purifying total nucleic acid by using gold-magnetic particles

Examples

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Embodiment 1

[0043] The method for purifying total nucleic acid with the gold magnetic particles of the present invention will be further described in conjunction with the following examples. Embodiment 1: The method for purifying total nucleic acid from rat liver tissue with gold magnetic particles

[0044] 1) Prepare sample lysate containing total nucleic acid

[0045] 1.1) Take 50 mg of rat liver tissue and put it into a glass homogenizer, add 1 ml of guanidinium salt lysate to homogenize. The guanidine salt lysate contains 2M guanidine hydrochloride;

[0046] 1.2) Transfer the sample lysate into a 2ml centrifuge tube and place it on ice for later use;

[0047] 2) combine

[0048] 2.1) Take 100 μl of gold magnetic particles in another 2ml centrifuge tube, place it on a magnetic separator for magnetic separation until the supernatant is clear, and discard the supernatant;

[0049]2.2) Remove the centrifuge tube, add 600 μl of binding buffer to the tube, mix well and set aside. The b...

Embodiment 2

[0055] Embodiment 2: the method for purifying total RNA from mouse liver tissue

[0056] 1) Prepare sample lysate containing total nucleic acid

[0057] 1.1) Take 50 mg of mouse liver tissue and put it into a glass homogenizer, add 1 ml of lithium salt lysate for homogenization. The lithium salt lysate contains 3M LiCl;

[0058] 1.2) Transfer the sample lysate into a 2ml centrifuge tube and place it on ice for later use;

[0059] 2) combine

[0060] 2.1) Take 100 μl of gold magnetic particles in another 2ml centrifuge tube, place it on a magnetic separator for magnetic separation until the supernatant is clear, and discard the supernatant;

[0061] 2.2) Remove the centrifuge tube, add 600 μl of binding buffer to the tube, mix well and set aside. The binding buffer contains 1M NaCl and 10% (W / V) polyethylene glycol, wherein the molecular weight of polyethylene glycol is 8000;

[0062] 2.3) Use a pipette to pipette 300 μl of the sample lysate from step 1.2) into the centrif...

Embodiment 3

[0073] Embodiment 3: the method for purifying total nucleic acid from rabbit blood

[0074] 1) Prepare sample lysate containing total nucleic acid

[0075] 1.1) Take 600 μl of fresh rabbit blood anticoagulated blood into a 2ml centrifuge tube, add 600 μl of guanidinium salt lysate and blow and mix. The guanidine salt lysate contains 2M guanidine hydrochloride;

[0076] 1.2) Centrifuge at 3000 g for 1 min, shake the precipitate to the bottom of the tube, absorb about 600 μl of supernatant and put it in another centrifuge tube for later use;

[0077] 2) combine

[0078] 2.1) Take 100 μl of gold magnetic particles in another 2ml centrifuge tube, place it on a magnetic separator for magnetic separation until the supernatant is clear, and discard the supernatant;

[0079] 2.2) Remove the centrifuge tube, add 600 μl of binding buffer to the tube, mix well and set aside. The binding buffer contains 1M NaCl and 10% (W / V) polyethylene glycol, wherein the molecular weight of polyeth...

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Abstract

The invention belongs to the field of nucleic acid purification, and in particular relates to a method for purifying total nucleic acid (comprising genome DNA and total RNA) by using gold-magnetic particles. The method comprises the following steps: preparing sample lysing solution containing the total nucleic acid; mixing the gold-magnetic particles and the biological sample lysing solution to form a gold-magnetic particle-total nucleic acid compound under the action of binding buffer solution; magnetically separating and cleaning the gold-magnetic particle-total nucleic acid compound; and finally, eluting the total nucleic acid from the surfaces of the gold-magnetic particles by using eluent, and collecting the supernate after magnetic separation to obtain purified total nucleic acid solution. According to the purifying method, centrifugation is saved, so the operation is simple and convenient, the purifying process is quick, the quality of the obtained total nucleic acid is high and the cost is low, and the genome DNA and the RNA in the biological sample can be obtained by separating at the same time.

Description

technical field [0001] The invention belongs to the field of nucleic acid purification, and relates to a method for purifying total nucleic acid (including genomic DNA and total RNA), in particular to a method for purifying total nucleic acid with gold magnetic particles. Background technique [0002] In the past few decades, molecular biology research has experienced rapid development. With the advancement of technology, nucleic acid purification has become an essential work in the preparation stage of molecular biology research. The first step in experimental operations such as disease diagnosis, cloning, sequencing, amplification, hybridization, and cDNA analysis is to purify complete and high-quality nucleic acids. Therefore, seeking a fast, reliable and reproducible nucleic acid purification method has always been of concern to people. [0003] The nucleic acid purification methods currently used can be roughly divided into two categories: liquid phase separation and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 崔亚丽张景阁安德鲁·G·陈刘蓓
Owner XIAN GOLDMAG NANOBIOTECH
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