Nucleic acid recovery and purification kit and method

A purification method and kit technology, applied in the biological field, can solve the problems of unfavorable sample preparation, heavy workload, and long elapsed time, and achieve the effect of shortening the time of sol, reducing surface tension, and simple and efficient components

Inactive Publication Date: 2015-10-14
SHANGHAI MAJORBIO BIO PHARM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing nucleic acid purification method for agarose gel recovery is cumbersome, takes a long time, has a large workload, and the range of recovered fragments is small, which is not conducive to the preparation of a large number of samples and is not conducive to the realization of high throughput.

Method used

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  • Nucleic acid recovery and purification kit and method
  • Nucleic acid recovery and purification kit and method

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Experimental program
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Effect test

Embodiment 1

[0031] The nucleic acid recovery and purification kit of this embodiment includes a sol solution, a magnetic bead solution and a binding solution. The sol contains 3.5 mol / L guanidine isothiocyanate and 25 mol / L Tris-Hcl buffer, and the rest is water. The pH value of the Tris-Hcl buffer used to configure the sol is 8. The magnetic bead solution contains 1 mg / mL of hydroxyl magnetic beads, and the rest is water. Hydroxyl magnetic beads were provided by Yingruicheng Biochemical Technology (Shanghai) Co., Ltd., the product model is 0501. Containing concentration is 50% isopropanol and the sodium chloride that concentration is 1.5mol / L in the binding solution, and the rest is water.

[0032] When the kit of this embodiment is used to recover and purify the nucleic acid of the agarose gel, the specific steps are as follows:

[0033] A. Use agarose gel electrophoresis to determine the position of the target DNA fragment in the agar block, then use a clean scalpel to cut off the a...

Embodiment 2

[0041] The nucleic acid recovery and purification kit of this embodiment includes a sol solution, a magnetic bead solution, and a binding solution. The sol solution contains guanidine isothiocyanate with a concentration of 3.2mol / L and a Tris-Hcl buffer solution with a concentration of 23mol / L, and the rest is water. The pH value of the Tris-Hcl buffer used to configure the sol is 8. The magnetic bead liquid contains 0.5mg / mL hydroxyl magnetic beads, and the rest is water. Containing concentration is 45% isopropanol and the sodium chloride that concentration is 1.2mol / L in the binding liquid, and the rest is water.

[0042] When the kit of this embodiment is used to recover and purify the nucleic acid of the agarose gel, the specific steps are as follows:

[0043] A. Use agarose gel electrophoresis to determine the position of the target DNA fragment in the agar block, then use a clean scalpel to cut off the agar block containing the target DNA fragment, and put it into a 1....

Embodiment 3

[0051] The nucleic acid recovery and purification kit of this embodiment includes a sol solution, a magnetic bead solution and a binding solution. The sol solution contains guanidine isothiocyanate with a concentration of 3.6 mol / L and a Tris-Hcl buffer solution with a concentration of 28 mol / L, and the rest is water. The pH value of the Tris-Hcl buffer used to configure the sol is 8. The magnetic bead liquid contains 2mg / mL hydroxyl magnetic beads, and the rest is water. Containing concentration is 55% isopropanol and the sodium chloride that concentration is 1.8mol / L in the binding solution, and the rest is water.

[0052] When the kit of this embodiment is used to recover and purify the nucleic acid of the agarose gel, the specific steps are as follows:

[0053] A. Use agarose gel electrophoresis to determine the position of the target DNA fragment in the agar block, then use a clean scalpel to cut off the agar block containing the target DNA fragment, and put it into a 1...

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Abstract

The invention relates to a nucleic acid recovery and purification kit which comprises a sol solution, a magnetic bead solution and a combination solution. The sol solution contains 3-4 mol/L guanidine isosulfocyanate, 20-30 mol/L Tris-HCl buffer solution and the balance of water. The magnetic bead solution contains 0.2-3 mg/mL hydroxy magnetic bead and the balance of water. The combination solution contains 40-60% isopropanol, 1-2 mol/L sodium chloride and the balance of water. The invention also relates to a nucleic acid recovery and purification method using the kit. The nucleic acid recovery and purification kit and method can quickly and efficiently recover and purify large-segment nucleic acids in a sepharose gel.

Description

technical field [0001] The invention relates to a kit for recovering and purifying nucleic acid and a method for using the kit, belonging to the field of biotechnology. Background technique [0002] Nucleic acid purification is an important prerequisite for downstream experiments in molecular biology and clinical detection of nucleic acid levels. The general principle of purification is to obtain high-purity nucleic acid samples and ensure the integrity of nucleic acid structures. Existing nucleic acid purification methods include phenol-chloroform extraction, high-salt precipitation, ion exchange, and selective adsorption. However, the method has disadvantages such as toxic reagents, cumbersome operation, inability to achieve high throughput, and limited size range of recovered fragments. Nano-micron superparamagnetic composite particles show great advantages in nucleic acid purification. Compared with the existing nucleic acid purification technology, it does not require ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 韩继臣李胜彬于丹徐亚楠
Owner SHANGHAI MAJORBIO BIO PHARM TECH
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