Kit for extracting plasma free DNA by paramagnetic particle method and use method of kit

A magnetic bead method and kit technology, applied in the field of DNA detection, can solve the problems of plasma free DNA loss, protein residue, and affecting the purity of plasma free DNA, and achieve the effects of long-term storage, effective extraction, and prevention of precipitation

Pending Publication Date: 2021-06-29
武汉远赞吉诺百客医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cell-free DNA extraction kits currently on the market have problems such as loss of plasma cell-free DNA and protein residues during the extraction process, which affects the purity of plasma cell-free DNA

Method used

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  • Kit for extracting plasma free DNA by paramagnetic particle method and use method of kit
  • Kit for extracting plasma free DNA by paramagnetic particle method and use method of kit
  • Kit for extracting plasma free DNA by paramagnetic particle method and use method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A kit for extracting plasma free DNA by the magnetic bead method, comprising the following components: lysate lyophilized powder, proteinase K, binding solution, magnetic bead solution, first cleaning solution, second cleaning solution and eluent, the above solution The solvent used was deionized water.

[0044]Lysis liquid freeze-dried powder: EDTA solution with a pH value of 8.0 and a concentration of 0.45mol / L, a sodium citrate solution with a concentration of 0.5mol / L, and a sodium alginate solution with a concentration of 0.1mol / L and a concentration of 0.25mol / L cetyltrimethylammonium bromide solution and Tween 20 with a volume fraction of 0.1%.

[0045] Binding solution: prepared from guanidine isothiocyanate solution with a concentration of 4 mol / L, sodium acetate solution with a concentration of 0.1 mol / L and triton x-100 solution with a volume fraction of 0.1%.

[0046] The preparation method of lysate freeze-dried powder is as follows: add 0.3% xanthan gum ...

Embodiment 2

[0062] A kit for extracting plasma free DNA by magnetic bead method, comprising the following components: lysate lyophilized powder, proteinase K, conjugated lyophilized powder, magnetic bead solution, first cleaning solution, second cleaning solution and eluent , the solvent of the above solution is deionized water.

[0063] Lysis solution freeze-dried powder: EDTA solution with a pH value of 8.0 and a concentration of 1mol / L, a sodium citrate solution with a concentration of 3mol / L, and a sodium alginate solution with a concentration of 1.3mol / L, and a concentration of 0.8mol / L cetyltrimethylammonium bromide solution and Tween 20 with a volume fraction of 2%.

[0064] Freeze-dried powder of binding solution: prepared from guanidine isothiocyanate solution with a concentration of 8 mol / L, sodium acetate solution with a concentration of 2 mol / L and triton x-100 solution with a volume fraction of 1%.

[0065] The preparation method of the lyophilized powder of the combined sol...

Embodiment 3

[0080] A kit for extracting plasma free DNA by magnetic bead method, comprising the following components: lysate lyophilized powder, proteinase K, conjugated lyophilized powder, magnetic bead solution, first cleaning solution, second cleaning solution and eluent , the solvent of the above solution is deionized water.

[0081] Lysis solution freeze-dried powder: EDTA solution with a pH value of 8.0 and a concentration of 0.5mol / L, a sodium citrate solution with a concentration of 1mol / L, and a sodium alginate solution with a concentration of 1mol / L and a concentration of 0.5mol / L cetyltrimethylammonium bromide solution and Tween 20 with a volume fraction of 0.5%.

[0082] Freeze-dried powder of binding solution: prepared from guanidine isothiocyanate solution with a concentration of 5 mol / L, sodium acetate solution with a concentration of 0.5 mol / L and triton x-100 solution with a volume fraction of 0.5%.

[0083] The preparation method of the lyophilized powder of the combine...

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Abstract

The invention provides a kit for extracting plasma free DNA by a paramagnetic particle method and a use method of the kit. The kit comprises a lysis solution, protease K, a binding solution, a magnetic particle solution, a first cleaning solution, a second cleaning solution and an eluent, wherein the lysis solution and the binding solution are concentrated to prepare lysate freeze-dried powder and binding solution freeze-dried powder respectively. According to the kit, through a specific reagent combination, repeated centrifugation is not needed in the use process, the total amount of plasma free DNA is guaranteed, cross contamination possibly caused in the centrifugation process is avoided, and the DNA purity is improved.

Description

technical field [0001] The invention relates to the field of DNA detection, in particular to a kit for extracting plasma free DNA by a magnetic bead method and a use method thereof. Background technique [0002] Plasma is the liquid component of blood except blood cells, white blood cells and platelets, accounting for about 50% of the whole blood volume. Studies have shown that plasma contains water, plasma proteins, sugars, and deoxyribonucleic acid molecules that are free outside the cells, which is plasma free DNA. Most of plasma cell-free DNA are double-stranded DNA molecules, whose fragment length is much smaller than that of genomic DNA, mainly concentrated between 0.1 and 0.3kb. Cell-free DNA analysis in plasma has potential applications throughout the natural course of cancer development, diagnosis, and treatment. In recent years, the application of plasma cell-free DNA in medical diagnosis has been continuously expanded, and it is widely used in non-invasive prena...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2523/308C12Q2527/125C12Q2521/537
Inventor 易吉解青青李泽卿
Owner 武汉远赞吉诺百客医学科技有限公司
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