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Extraction method of clotting genomic DNA (deoxyribonucleic acid)

An extraction method and genomic technology, applied in the field of genomic DNA extraction, can solve the problems of unsatisfactory DNA yield and purity, achieve the effect of increasing purity and yield, and improving extraction speed

Inactive Publication Date: 2012-01-18
吴剑 +1
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  • Abstract
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Problems solved by technology

[0004] The object of the present invention is to provide a method for extracting blood coagulation genomic DNA, so as to solve the problem that the DNA yield and purity are not ideal when extracting genomic DNA from whole blood containing blood clots

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  • Extraction method of clotting genomic DNA (deoxyribonucleic acid)
  • Extraction method of clotting genomic DNA (deoxyribonucleic acid)
  • Extraction method of clotting genomic DNA (deoxyribonucleic acid)

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Embodiment 1

[0033] Material source: nattokinase, 40000Fu / g, purchased from Shanghai Weida Company; proteinase K was sigma proteinase K.

[0034] Sample source: Sample 1: Fresh blood clot (with serum) rabbit blood; Sample 2: Fresh blood clot (without serum) rabbit blood; Sample 3: Human blood clot stored at -20°C for one month; Sample 4: -20 ℃ to preserve human blood clots for a week. The sample collection volume is 250 μL, and the following processing is carried out:

[0035] 1. Transfer the sample to a 1.5mL centrifuge tube. If the sample volume is less than 250ul, add PBS or TE Buffer to make up to 250μL, add 20mg nattokinase, and let it stand at room temperature for 5min until the clot disappears; add 25μL proteinase K and 250μL proteinase K to the centrifuge tube Lysis solution, shake for 10 seconds to mix, add 5 μL RNase A; (lysate formula: 20% SDS, 1% Triton-x100, 100mM sodium acetate, 50mM EDTA, pH5.2)

[0036] 2. Incubate at 65°C for 10 minutes, shake and mix once during the per...

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Abstract

The invention provides an extraction method of clotting genomic DNA (deoxyribonucleic acid), which comprises the following steps that: nattokinase is added to a sample before extraction, and fibrin can be quickly decomposed by utilizing the thrombolytic force of the nattokinase several times more than that of a thrombolytic agent UK (urokinase), and further blood clots are separated, so that the blood clots become a homogeneous solution; and the obstacle of blood DNA extraction caused by sludged blood is removed, thereby the extraction speed of blood DNA is obviously improved, and simultaneously the purity and the yield of DNA product are greatly increased. The method provided by the invention is applicable to the extraction of DNA from sludged blood from a crime scene or dead bodies, the extraction of high-throughput blood gDNA and the extraction of blood gDNA in an antibody analysis experiment.

Description

technical field [0001] The invention relates to a method for extracting genome DNA, in particular to a method for extracting genome DNA from human blood clots. Background technique [0002] DNA extracted from blood can be widely used in disease diagnosis, genetic analysis, identification, etc. Medicine is developing rapidly from traditional medical treatment to molecular biological test diagnosis and auxiliary treatment. In the process of blood DNA extraction, especially in the process of high-throughput blood DNA extraction, due to various reasons, blood clots often occur in individual samples. In addition, residual blood samples from crime scenes can also produce a large number of clots due to prolonged storage. The resulting clot will hinder the smooth progress of high-throughput blood extraction. The current common blood DNA extraction requires the use of mechanical grinding to grind the clot and then extract the blood DNA, which is time-consuming, inconvenient to ope...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 吴剑匡虹桥
Owner 吴剑
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