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Kit for rapidly extracting bacterial nucleic acid based on magnetic beads and extraction method

A bacterial nucleic acid and kit technology, applied in the biological field, can solve the problems of complex extraction steps of bacterial nucleic acid, low extraction efficiency, and many types of reagents, and achieve the effects of low toxicity, high nucleic acid purity, and easy operation

Inactive Publication Date: 2021-04-20
GENFINE BIOTECH BEIJING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For this reason, the present invention provides a kind of test kit based on magnetic bead to extract bacterial nucleic acid quickly, to solve the problems such as complex step of extracting bacterial nucleic acid in the prior art, many types of reagents, toxic reagents such as phenol chloroform, low extraction efficiency, etc.

Method used

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  • Kit for rapidly extracting bacterial nucleic acid based on magnetic beads and extraction method
  • Kit for rapidly extracting bacterial nucleic acid based on magnetic beads and extraction method
  • Kit for rapidly extracting bacterial nucleic acid based on magnetic beads and extraction method

Examples

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Embodiment 1

[0030] Embodiment 1, utilize the kit of rapid extraction bacterial nucleic acid based on magnetic bead to manually extract bacterial nucleic acid method

[0031] 1.1 In this embodiment, the specific reagents in the kit for manually extracting nucleic acid using a kit for rapidly extracting bacterial nucleic acid based on magnetic beads include:

[0032] 1) Lysis solution: 5mol / L guanidine hydrochloride, 10% sodium lauryl sulfate, 50mmol / L ethylenediaminetetraacetic acid, 100mmol / L tris, 200mmol / L sodium chloride, 2% fatty alcohol Sodium polyoxyethylene ether sulfate, 10% TritonX-100, 5% PEG4000;

[0033] 2) Protein-removing rinse solution: 2mol / L guanidine salt, 500mmol / L sodium chloride, 80% ethanol (V / V);

[0034] 3) Desalting rinse solution: 80% ethanol (V / V);

[0035] 4) Nucleic acid eluent: 20mmol / L Tris.

[0036] 1.2 A kind of manual method utilizes the method for extracting bacterial nucleic acid based on the test kit of rapidly extracting bacterial nucleic acid base...

Embodiment 2

[0043] Embodiment 2, the method for automatically extracting bacterial nucleic acid using a kit for rapidly extracting bacterial nucleic acid based on magnetic beads

[0044] 2.1 The reagents in the kit for rapidly extracting bacterial nucleic acid based on magnetic beads used in this embodiment are the reagents in the kit described in Example 1;

[0045] Automatic nucleic acid extractor: Jifan Purifier32, including pre-splitting plates.

[0046] 2.2 A method for automatically extracting bacterial nucleic acid using a kit for rapidly extracting bacterial nucleic acid based on magnetic beads, comprising the following steps:

[0047] 1) Add the reagents in the kit to the pre-separation plate, add 500 μL deproteinized rinse solution to columns 1 and 7, add 300 μL lysate to columns 2 and 8, add 800 μL desalted rinse solution to columns 3 and 9, add 500 μL desalted rinse solution to columns 6 and 12 Add 100 μL of nucleic acid eluent to the column;

[0048] 2) Add 200 μL of the cl...

experiment example 1

[0051] Experimental Example 1 The method of manually extracting Escherichia coli sample nucleic acid using a kit for rapidly extracting bacterial nucleic acid based on magnetic beads

[0052] 1.1 In this experimental example, a kit for rapid extraction of bacterial nucleic acid based on magnetic beads is used to manually extract nucleic acid from Escherichia coli samples. The specific reagents in the kit include:

[0053] 1) Lysis solution: 5mol / L guanidine hydrochloride, 10% sodium lauryl sulfate, 50mmol / L ethylenediaminetetraacetic acid, 100mmol / L tris, 200mmol / L sodium chloride, 2% fatty alcohol Sodium polyoxyethylene ether sulfate, 10% TritonX-100, 5% PEG4000;

[0054] 2) Protein-removing rinse solution: 2mol / L guanidine salt, 500mmol / L sodium chloride, 80% ethanol (V / V);

[0055] 3) Desalting rinse solution: 80% ethanol (V / V);

[0056] 4) Nucleic acid eluent: 20mmol / L Tris.

[0057] 1.2 Method for manual extraction of nucleic acid from Escherichia coli samples using a ma...

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Abstract

The invention discloses a kit for rapidly extracting the bacterial nucleic acid based on magnetic beads and an extraction method. Reagents of the kit include a lysis solution, a magnetic bead solution, a deproteinization rinsing solution, a desalination rinsing solution and a nucleic acid eluent. The method comprises the steps of adding the magnetic bead solution and the lysis solution into a sample for lysis, performing rinsing with the rinsing solutions, and performing eluting with the nucleic acid eluent to obtain the bacterial nucleic acid. The kit for rapidly extracting bacterial nucleic acid based on the magnetic beads according to the invention is simple and convenient to extract, does not contain toxic reagents such as phenol chloroform, can be manually operated, can be integrated with various automatic nucleic acid extractors, has simple and convenient steps, and can automatically extract the nucleic acid only by adding a sample and without adding protease K, simplifies experimental operation steps to the greatest extent and guarantees the safety of an experimenter. The requirements on the nucleic acid extraction speed and flux and the requirements on the nucleic acid extraction quality when more clinical samples exist in a hospital are met; and meanwhile, the kit according to the invention is low in toxicity, simple and convenient to operate, high in sample flux and better in nucleic acid extraction effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for rapidly extracting bacterial nucleic acid based on a magnetic bead method and a method for extracting bacterial nucleic acid. Background technique [0002] At present, there are many methods or kits for bacterial genome extraction on the market. The main methods include traditional heating and boiling method, Chelex-100 extraction method, SDS-NaOH method, SDS-enzyme cleavage method, CTAB method, kit spin column method . [0003] However, these methods currently have some problems, including complicated steps, many types of reagents, toxic reagents such as phenol and chloroform, and low extraction efficiency. Contents of the invention [0004] For this reason, the present invention provides a kit for rapidly extracting bacterial nucleic acid based on magnetic beads to solve the problems in the prior art that the extraction of bacterial nucleic acid is complicated, there a...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 韩典霖胡美丽杨亮
Owner GENFINE BIOTECH BEIJING CO LTD
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