Nucleic acid releaser and method for quickly extracting nucleic acids from dried blood spots

The technology of a nucleic acid releasing agent and an extraction method is applied in the field of nucleic acid releasing agent and fast extraction of nucleic acid from dried blood spots, which can solve the problems such as inability to effectively remove impurities such as hemoglobin, and achieve the effects of low cost, easy identification and convenient operation.

Inactive Publication Date: 2016-05-25
王智勇
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The technical problem to be solved by the present invention is that the current nucleic acid release method of dried blood spots mainly has the problem of not being able to effectively remove impurities such as hemoglobin

Method used

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  • Nucleic acid releaser and method for quickly extracting nucleic acids from dried blood spots
  • Nucleic acid releaser and method for quickly extracting nucleic acids from dried blood spots
  • Nucleic acid releaser and method for quickly extracting nucleic acids from dried blood spots

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Experimental program
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Effect test

Embodiment 1

[0073] Preparation of nucleic acid release agent: Tris-HCl, EDTA, MgCl with a pH value of 7.4 2 , TrixonX-100, KCL and proteinase K were added into sterile water and mixed evenly, among which Tris-HCl, EDTA, MgCl 2 , TrixonX-100, KCL, and Proteinase K are at concentrations of 13mM / L, 2mM / L, 14.5mM / L, 2mM / L, 10mM / L, and 0.065mg / mL, and are stored at -20°C;

[0074] Preparation of blood spot sample discs: Separate a 5×5mm sample disc from the dried blood spot collection card, put it into a centrifuge tube, and use a puncher to punch a 5×5mm blood spot (or a round A blood spot cut into a sample of approximately 5×5 mm).

[0075] The nucleic acid release solution was transferred to a filter column, and centrifuged at a centrifugal force of 5000×g for 1 min to obtain a filtrate.

[0076] Transfer the filtrate to another clean, enzyme-free centrifuge tube and store it at -20°C to obtain the finally extracted and purified dried blood spot nucleic acid release agent. The parameters ...

Embodiment 2

[0091] 1L solution of nucleic acid release agent, in which Tris-HCl, EDTA, MgCl 2 , TrixonX-100, KCL and proteinase K concentrations were 10mM / L, 2.5mM / L, 10mM / L, 5mM / L, 14.005mM / L, 0.06mg / mL.

Embodiment 3

[0093] 1L solution of nucleic acid release agent, in which Tris-HCl, EDTA, MgCl 2 , TrixonX-100, KCL and proteinase K concentrations were 20mM / L, 1mM / L, 12mM / L, 1mM / L, 7.485mM / L, 0.08mg / mL.

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Abstract

The invention discloses a nucleic acid releaser and a method for quickly extracting nucleic acids from dried blood spots. The nucleic acid releaser is formed by adding Tris-HCl, EDTA (ethylene diamine tetraacetic acid), MgCl2, Trixon X-100, KCl and proteinase K into sterilized water and mixing. The concentration of the Tris-HCl is 10-50 mM/L, the concentration of the EDTA is 1-5 mM/L, the concentration of the MgCl2 is 10-50 mM/L, the concentration of the Trixon X-100 is 1-10 mM/L, the concentration of the KCl is 2-20 mM/L, and the concentration of the proteinase K is 0.05-0.1 mg/mL. The method for quickly extracting nucleic acids from dried blood spots utilizes the nucleic acid releaser to obtain the target nucleic acids. The nucleic acid releaser and method are convenient to operate, can be used for quickly extracting and purifying nucleic acids in dried blood spots and other blood samples, and have the advantages of high repetitiveness, high specificity, lower cost, higher use safety and wide applicability.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nucleic acid releasing agent and a method for rapidly extracting dried blood spot nucleic acid. Background technique [0002] Dried blood spot (DBS), a blood collection method that collects whole blood samples on cardboard, has certain advantages over traditional methods. It requires less blood volume and is convenient for blood sample collection, storage and transportation. It is a rare disease screening method. An effective tool in the investigation process. Dried blood spots have been used in newborn screening for phenylketonuria since the 1960s. With the development of tandem mass spectrometry technology, the application value of dried blood spots in the diagnosis and screening of genetic metabolic diseases has become increasingly prominent. As of May 2015, dried blood spots are mainly used in the screening of metabolic-related diseases, pharmacokinetic studies, and newborn s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1017C12Q2521/537C12Q2527/125
Inventor 王智勇
Owner 王智勇
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