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Kit for rapidly extracting free nucleic acid in plasma based on paramagnetic particle method

A free nucleic acid and kit technology, applied in the field of biomedicine, can solve the problems of inability to achieve high-throughput nucleic acid extraction, high toxicity of reagents, and low nucleic acid yield, and achieve the effects of low toxicity, simple steps and high nucleic acid quality.

Active Publication Date: 2022-02-18
GENFINE BIOTECH BEIJING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] Traditional free nucleic acid extraction kits usually use precipitation method and column membrane centrifugation method. The precipitation method needs to use phenol chloroform extraction, which is complicated to operate and highly toxic reagents, so it is not suitable for use
The column membrane method needs to be used with a centrifuge, the operation is complicated, the nucleic acid yield is low, and high-throughput nucleic acid extraction cannot be achieved

Method used

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  • Kit for rapidly extracting free nucleic acid in plasma based on paramagnetic particle method
  • Kit for rapidly extracting free nucleic acid in plasma based on paramagnetic particle method
  • Kit for rapidly extracting free nucleic acid in plasma based on paramagnetic particle method

Examples

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Embodiment 1

[0055] Embodiment 1, utilize the kit of the magnetic bead method of the present invention to extract plasma free nucleic acid to manually extract plasma free nucleic acid

[0056] 1.1 In this embodiment, the specific reagents in the kit for manually extracting nucleic acid using a kit for rapidly extracting free plasma nucleic acid based on magnetic beads include:

[0057] 1) Lysis solution: 3mol / L guanidine isothiocyanate, 2% sodium lauryl sulfate, 100mmol / L ethylenediaminetetraacetic acid, 100mmol / L tris, 100mmol / L sodium chloride, 2 % sodium dodecylbenzene sulfonate, 2.5% sodium laureth sulfate, 8% TritonX-100;

[0058] 2) Protein removal rinse solution: 2mol / L guanidine isothiocyanate, 500mmol / L sodium chloride, 80% ethanol (V / V);

[0059] 3) Desalting rinse solution: 80% ethanol (V / V);

[0060] 4) Nucleic acid eluent: 10mmol / L Tris, pH 8.0.

[0061] 1.2 A manual method utilizing a magnetic bead-based rapid plasma free nucleic acid extraction kit to extract plasma free ...

Embodiment 2

[0069] With reference to Example 1 preparation reagent, wherein lysate contains 1% sodium dodecylbenzene sulfonate, 3.5% sodium lauryl ether sulfate, other compositions are identical;

[0070] Referring to the operation of 1.2 in Example 1, the obtained nucleic acid yield and DNA library construction yield are shown in Table 2.

Embodiment 3

[0072] With reference to Example 1 preparation reagent, wherein lysate contains 3% sodium dodecylbenzene sulfonate, 1.5% sodium lauryl ether sulfate, other compositions are identical;

[0073] Referring to the operation of 1.2 in Example 1, the obtained nucleic acid yield and DNA library construction yield are shown in Table 2.

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Abstract

The invention discloses a kit for rapidly extracting free nucleic acid from plasma based on a paramagnetic particle method. The kit comprises a lysate, a deproteinization rinsing solution, a desalting rinsing solution, an eluent and magnetic beads, wherein the lysate contains a chaotropic agent with a concentration of 2 to 6 M, 0.05 to 10% of lauryl sodium sulfate, ethylenediamine tetraacetic acid with a concentration of 50 to 200 mM, trihydroxymethyl aminomethane with a concentration of 10 to 200 mM, sodium chloride with a concentration of 50 to 500 mM, 0.1 to 5% of sodium dodecyl benzene sulfonate, 1 to 5% of sodium laureth sulfate and 5 to 15% of a nonionic surfactant. According to the method, unique lysate components and the magnetic beads are adopted, protease K does not need to be added, nucleic acid yield can be effectively increased, and therefore, the yield of downstream NGS library building is increased.

Description

technical field [0001] The invention provides a kit for rapidly extracting plasma free nucleic acid based on a magnetic bead method, which belongs to the field of biomedicine. Background technique [0002] Plasma free nucleic acid refers to the partially degraded endogenous DNA that exists in human plasma and is free outside the cells. Cell-free nucleic acid detection is widely used in non-invasive prenatal screening, tumor diagnosis and various disease testing. [0003] At present, there are many methods for extracting free nucleic acid from plasma in the market. According to the principle of nucleic acid extraction, it can be divided into precipitation method, column membrane centrifugation method, magnetic bead method, etc., and can be divided into manual method and instrument method according to the extraction flux. [0004] Precipitation method: Extract nucleic acid with organic reagents such as phenol chloroform. [0005] Column membrane centrifugation: DNA is bound ...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/6806
CPCC12N15/1013C12Q1/6806C12Q2527/125C12Q2523/308
Inventor 胡美丽韩典霖俞萍
Owner GENFINE BIOTECH BEIJING CO LTD
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