Genome magnetic bead extraction test kit and extraction method

A kit and genome technology, applied in the field of genomic magnetic bead extraction kits, can solve the problems of low extraction nucleic acid concentration, limited application of magnetic bead method, low extraction efficiency, etc. good purity

Inactive Publication Date: 2020-11-20
华芯生物科技(武汉)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, mucin and mucopolysaccharides are the main substances that make saliva sticky, which limits the application of magnetic bead method in the extraction of saliva nucleic acid.
[0005] The existing magnetic bead extraction kits have the following disadvantages: low extraction efficiency, low concentration of extracted nucleic acid, and poor purity

Method used

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  • Genome magnetic bead extraction test kit and extraction method
  • Genome magnetic bead extraction test kit and extraction method
  • Genome magnetic bead extraction test kit and extraction method

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Embodiment approach

[0040] According to a typical embodiment of the present invention, a genome magnetic bead extraction kit is provided, the kit comprising: lysate, proteinase K, isopropanol, hydroxyl magnetic beads, washing solution and eluent;

[0041] The formula of the lysate is: 25-75mM Tris-HCL with pH 7.5-8.5, 3-5M guanidine isothiocyanate, 5-15% polidocanol, 5-15% PEG8000;

[0042] The washing solution includes A liquid and B liquid, the A liquid is: 120-130mmol / l NaCl, the volume concentration is 80-90% dehydrated alcohol; the B liquid is: 25-35mmol / l Tris, the volume concentration is 80% ~90% absolute ethanol;

[0043] The formula of the eluent is: 9.5-10.5mM Tris-HCl; 0.8-1.2mM EDTA.

[0044] The invention provides a genome magnetic bead extraction kit, the nucleic acid concentration is higher, and the nucleic acid agarose gel electrophoresis has no miscellaneous bands and dragging phenomena, indicating that the purity of the extracted nucleic acid is better; the lysate does not need...

Embodiment 1

[0073] 1. A genomic magnetic bead extraction kit provided in an embodiment of the present invention, comprising: lysate, proteinase K, isopropanol, washing liquid, eluent, and hydroxyl magnetic beads;

[0074] The formula of the lysate is: 50mM Tris-HCL with pH 8.0, 4M guanidine isothiocyanate, 10% polidocanol, 10% PEG8000;

[0075] The washing solution includes A liquid and B liquid, the A liquid is: 125mmol / l NaCl, the volume concentration is 85% dehydrated alcohol; the B liquid is: 30mmol / l Tris, the volume concentration is 85% dehydrated alcohol;

[0076] The formula of the eluent is: 8mM Tris-HCl; 0.5mM EDTA.

[0077] 2. How to use Genome Magnetic Beads Extraction Kit

[0078] (1) Sample Lysis and Combination:

[0079] Add 125 μL sample, 20 μL proteinase K solution, and 500 μL lysate to a 1.5 mL centrifuge tube, add 350 μL isopropanol to the above centrifuge tube, then add 20 μL magnetic bead suspension, vortex (speed 9-10) ) for 3 minutes and then allowed to stand for...

Embodiment 2

[0092] 1. A genomic magnetic bead extraction kit provided in an embodiment of the present invention, comprising: lysate, proteinase K, isopropanol, washing liquid, eluent, and hydroxyl magnetic beads;

[0093] The formula of the lysate is: 25mM Tris-HCL with pH7.5, 3M guanidine isothiocyanate, 5% polidocanol, 5% PEG8000;

[0094] The washing liquid includes A liquid and B liquid, the A liquid is: 120mmol / l NaCl, the volume concentration is 80% dehydrated ethanol; the described B liquid is: 25mmol / l Tris, the volume concentration is 80% dehydrated ethanol;

[0095] The formula of the eluent is: 9.5mM Tris-HCl; 0.8mM EDTA.

[0096] 2. The method of using the Genome Magnetic Beads Extraction Kit is the same as in Example 1. Figure 7 It is the target gene detection and nucleic acid agarose gel electrophoresis image in Example 1; the target gene detection band is brighter, and it is found that the nucleic acid has no entrainment and dispersion phenomenon by running the gel on the...

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Abstract

The invention discloses a genome magnetic bead extraction test kit. The test kit comprises a lysis solution, proteinase K, isopropanol, hydroxyl magnetic beads, a washing solution and an eluent, wherein the formula of the lysis solution comprises 25 to 75 mM of Tris-HCL with a pH value of 7.5 to 8.5, 3 to 5 M of guanidine isothiocyanate, 5 to 15% of polidocanol and 5 to 15% of PEG8000; the washingsolution comprises a solution A and a solution B; the solution A comprises NaCl with a concentration of 120 to 130 mmol / l and absolute ethyl alcohol with a volume concentration of 80 to 90%; the solution B comprises Tris with a concentration of 25 to 35 mmol / l and absolute ethyl alcohol with a volume concentration of 80 to 90%; and the formula of the eluent comprises 9.5 to 10.5 mM of Tris-HCl and 0.8 to 1.2 mM of EDTA. The concentration of the nucleic acid extracted by the test kit is higher; nucleic acid agarose gel electrophoresis is free of the phenomenons of miscellaneous bands and bandtrailing; and the purity of extracted nucleic acid is better.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a genome magnetic bead extraction kit and an extraction method. Background technique [0002] Nucleic acid extraction methods include phenol-chloroform method, salting-out method and column adsorption method. Traditional methods for nucleic acid extraction have their own advantages and disadvantages, but the extraction effect is not good, and it is only suitable for manual extraction, not for automation. The magnetic bead method utilizes the specific adsorption of magnetic beads to nucleic acid, separates and obtains nucleic acid under the action of a magnetic field, and is suitable for automatic extraction. [0003] Magnetic beads have good surface effect and volume effect, good selective magnetic response, stable physical and chemical properties, etc., which make the bead method nucleic acid extraction widely used. However, different magnetic bead nucleic acid extraction methods ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 杨思雨郭静董思远张雪娇赵愿安舒芹
Owner 华芯生物科技(武汉)有限公司
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