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Amino acid buffer solution and plasma free DNA extraction kit

A technology of amino acids and buffers, applied in the field of biomedicine, to achieve the effect of ensuring integrity, low cost, and promoting capture efficiency

Active Publication Date: 2019-08-16
合肥欧创基因生物科技有限公司
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  • Application Information

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Problems solved by technology

Studies have shown that amino acid buffers are as effective as high-concentration chaotropic salts, but there is no clear literature on the role of amino acids in combination with low-concentration chaotropic salts in promoting nucleic acid adsorption

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  • Amino acid buffer solution and plasma free DNA extraction kit
  • Amino acid buffer solution and plasma free DNA extraction kit
  • Amino acid buffer solution and plasma free DNA extraction kit

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Embodiment Construction

[0032] The present invention will be specifically introduced below in conjunction with the accompanying drawings and specific embodiments.

[0033] An amino acid buffer, including: 80-200mM amino acid, 0.1-0.3M chaotropic salt, 5%-15% PEG, 0.5-1.2M sodium chloride, 40-100mM Tris-HCl, 20-50mM EDTA, pH: 2.1-2.4.

[0034] A plasma cell-free DNA extraction kit, comprising the following reagents:

[0035] Lysis solution, proteinase K working solution, amino acid adsorption buffer, washing solution, TE eluent;

[0036] The lysate includes: 20-100mM Tris-HCl, 50-150mM NaCl, 5-10% glycerol, 1-10% SDS, 1-2mM EDTA, pH 7.0-8.0;

[0037] Proteinase K working solution includes: Dissolve 20mg of proteinase K in 1ml of proteinase K buffer to obtain a working solution with a concentration of 20mg / ml; proteinase K buffer: 0.2-2% SDS, pH7.5-8.0;

[0038] Amino acid adsorption buffer includes: 80-200mM amino acid, 0.1-0.3M chaotropic salt, 5%-15% PEG, 0.5-1.2M sodium chloride, 40-100mM Tris-H...

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Abstract

The invention discloses an amino acid buffer solution and a plasma free DNA extraction kit. The plasma free DNA extraction kit comprises the agents of a lysis solution, a proteinase K working liquid,an amino acid adsorption buffer solution, a washing solution, and a TE eluent; the amino acid adsorption buffer solution comprises 80-200 mM of amino acids, 0.1-0.3 M of a chaotropic salt, 5-15% of PEG (polyethylene glycol), 0.5-1.2 M of sodium chloride, 40-100 mM of Tris-HCl, and 20-50 mM of EDTA (ethylenediaminetetraacetic acid), and has pH of 2.1-2.4. the amino acids are combined with the chaotropic salt having a low concentration; pH of the buffer solution is controlled; therefore, extraction efficiency is improved. The use of PEG helps reduce shear injury of DNA and ensure DNA integrity.Therefore, extraction of free DNAs in plasma is free of inhibitor residue, has high quality, is simple to perform, and has low cost.

Description

technical field [0001] The invention relates to the technical field of biomedicine and biological sample pretreatment, in particular to an amino acid buffer solution and a plasma free DNA extraction kit. Background technique [0002] cfDNA is a fragmented extracellular DNA, mainly present in body fluids such as plasma, serum, and cerebrospinal fluid, with a length ranging from tens to hundreds of bases. At present, the main application areas of cfDNA are non-invasive prenatal diagnosis and tumor liquid biopsy. With the increasing application of cfDNA, the extraction of cfDNA is very important. The extraction of cfDNA includes the separation of nucleic acid and protein, adsorption of nucleic acid, removal of impurities such as protein, and elution of nucleic acid. [0003] At present, cfDNA extraction mostly uses phenol-chloroform method, magnetic bead method and silica gel membrane adsorption column method. Reagents such as phenol-chloroform are harmful to the human body, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q2527/125
Inventor 杨芳梅张惠贤卢德景徐红梅
Owner 合肥欧创基因生物科技有限公司
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