Kit for genome DNA extraction and application of kit

A kit and genome technology, applied in the field of genomic DNA extraction kits, can solve the problems of low yield, turbidity, low purity, etc., and achieve the effect of simple process, improved quality and efficiency

Inactive Publication Date: 2020-03-31
SHENZHEN HAPLOX BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extraction of genomic DNA from blood cells is prone to problems such as eluent viscosity and low purity
The extraction of genomic DNA from saliva is also prone to problems such as low yield, viscous eluate, and turbidity
[0006] In general, most of the genomic DNA extraction kits currently avai...

Method used

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  • Kit for genome DNA extraction and application of kit
  • Kit for genome DNA extraction and application of kit
  • Kit for genome DNA extraction and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The kit in this example specifically includes lysate, proteinase K, magnetic bead suspension, erythrocyte lysate, binding solution, cleaning solution 1, cleaning solution 2 and eluent.

[0045] Wherein, the lysate is composed of 100mmol / L Tris-HCl, 20mmol / L EDTA, 1.4mol / L NaCl, 1% PEG8000 and 1% Tween-20.

[0046] Proteinase K was purchased from Tiangen Biotechnology, and the magnetic bead suspension was purchased from Meiji Biotechnology.

[0047] Red blood cell lysate was composed of 30mmol / L Tris-HCl, 5mmol / L EDTA, 60mmol / L NaCl and 0.015% TritonX-100.

[0048] The binding solution is isopropanol.

[0049] The cleaning solution 1 is composed of 2mol / L guanidine hydrochloride and 50% absolute ethanol.

[0050] Cleaning solution 2 is composed of 50mmol / L NaCl and 75% absolute ethanol.

[0051] The eluent was nuclease-free sterile deionized water.

[0052] The method of using the kit in this example is as follows:

[0053] (1) For human tissue samples, take 30 mg o...

Embodiment 2

[0066] In this example, on the basis of Example 1, the components and concentrations of the components of the lysate, erythrocyte lysate, cleaning solution 1 and cleaning solution 2 were tested, as detailed below:

[0067] 1. Lysate formulation test

[0068] In this example, different concentrations of the components in the lysate were tested, as shown in Table 2.

[0069] Table 2 Lysate formula test

[0070]

[0071]

[0072] In this example, lysates of seven formulations from Test 1 to Test 7 were prepared according to Table 2, and the rest of the reagents, such as proteinase K, erythrocyte lysate, binding solution, cleaning solution 1 and cleaning solution 2, etc., were the same as those in Example 1. In the same method as in Example 1, genomic DNA was extracted from the two human tissue samples, the two blood cell samples and the two saliva samples in Example 1. And measure the concentration and purity of the extracted genomic DNA according to the method of Example...

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Abstract

The invention discloses a kit for genome DNA extraction and an application of the kit. The kit comprises a lysis solution and an erythrocyte lysis solution, wherein the lysis solution contains 20 to 300mM of Tris-HCl, 20 to 100mM of EDTA, 0.3 to 2M of NaCl, 0.5 to 5 percent of PEG8000 and 0.5 to 10 percent of Triton X-100, and SDS or Tween 20; the erythrocyte lysis solution contains 10 to 100mM ofTris-HCl, 1 to 50mM of EDTA, 10 to 300mM of NaCl and 0.01 to 0.2 percent of Triton X-100. According to the kit, through optimization of the lysis solution and the erythrocyte lysis solution, polysaccharide can be effectively removed, phenol is combined, phenol is prevented from being combined with DNA, the problems of viscosity and turbidity are solved, the kit is suitable for various different samples, and the purity and yield of extracted genome DNA can meet various downstream experiment use requirements.

Description

technical field [0001] This application relates to the technical field of genomic DNA extraction, in particular to a kit for genomic DNA extraction and its application. Background technique [0002] Nucleic acid is a very important class of biological macromolecules. It plays an irreplaceable role in a series of life processes such as reproduction, growth, development, aging and death of organisms. It can be called the "master" of the biological world. Now it has It has become an important research topic in many life science fields such as biochemistry, molecular biology, genetics, and life medicine. Therefore, extracting high-quality and high-purity nucleic acids from numerous and complex biological macromolecules is an essential first step for other research. [0003] The principles of DNA extraction include: ensuring the integrity of the primary structure of DNA; there should be no organic solvents that inhibit enzymes and excessively high concentrations of metal ions; o...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2527/125
Inventor 许明炎张晓妮沈广强张生
Owner SHENZHEN HAPLOX BIOTECH
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