Method for safely and quickly extracting genomic deoxyribose nucleic acid (DNA) from blood

A genome and blood technology, applied in the field of animal molecular biology technology research, to achieve the effects of good DNA quality, inhibition of degradation and simple operation

Inactive Publication Date: 2012-07-25
邢秀梅
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, although there are related kits, most of them contain toxic and harmful substances that are harmful to the body

Method used

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  • Method for safely and quickly extracting genomic deoxyribose nucleic acid (DNA) from blood
  • Method for safely and quickly extracting genomic deoxyribose nucleic acid (DNA) from blood
  • Method for safely and quickly extracting genomic deoxyribose nucleic acid (DNA) from blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Preparation of reagents:

[0035] (1) Low-salt buffer (pH 7.6): Dissolve 1.21g Tris base (trishydroxymethylaminomethane) and 3.38g Na in 800ml distilled water 2 EDTA (disodium ethylenediaminetetraacetic acid), 0.76g MgCl 2 , 0.47gNaCl. Add concentrated hydrochloric acid to adjust the pH to the desired value, add distilled water to make up to 1L.

[0036] (2) Cell lysate: Add 1g SDS (sodium dodecyl sulfate) and 1ml Triton X-100 (polyethylene glycol p-isooctylphenyl ether) to 200ml low-salt buffer.

[0037] Steps:

[0038] 1) Add 200 μl of anticoagulated blood and 400 μl of low-salt buffer solution from sika deer into a 1.5 ml centrifuge tube, mix thoroughly, centrifuge at 1 000 RPM for 10 min at room temperature, and discard the supernatant;

[0039] 2) Add 500 μl low-salt buffer, mix thoroughly, centrifuge at 1 000 RPM for 5 min at room temperature, and repeat the previous step;

[0040] 3) Carefully aspirate the supernatant, add 300 μl cell lysate, mix by pipettin...

Embodiment 2

[0046] Reagent preparation:

[0047] (1) Low-salt buffer (pH 7.6): Dissolve 1.21g Tris base (trishydroxymethylaminomethane) and 3.38g Na in 800ml distilled water 2 EDTA (disodium ethylenediaminetetraacetic acid), 0.76g MgCl 2 , 0.47gNaCl. Add concentrated hydrochloric acid to adjust the pH to the desired value, add distilled water to make up to 1L.

[0048] (2) Cell lysate: Add 1g SDS (sodium dodecyl sulfate) and 1ml Triton X-100 (polyethylene glycol p-isooctylphenyl ether) to 200ml low-salt buffer.

[0049] Steps:

[0050]1) Add 200 μl anticoagulant blood and 400 μl low-salt buffer solution from red deer to a 1.5 ml centrifuge tube, mix thoroughly, centrifuge at 1 000 RPM for 10 min at room temperature, and discard the supernatant;

[0051] 2) Add 500 μl low-salt buffer, mix thoroughly, centrifuge at 1 000 RPM for 5 min at room temperature, and repeat the previous step;

[0052] 3) Carefully aspirate the supernatant, add 300 μl cell lysate, mix by pipetting, and bath...

Embodiment 3

[0058] Reagent preparation:

[0059] (1) Low-salt buffer (pH 7.7): Dissolve 1.27g Tris base (trishydroxymethylaminomethane) and 3.5g Na in 800ml distilled water 2 EDTA (disodium ethylenediaminetetraacetic acid), 0.78g MgCl 2 , 0.46gNaCl. Add concentrated hydrochloric acid to adjust the pH to the desired value, add distilled water to make up to 1L.

[0060] (2) Cell lysate: Add 0.8 g of SDS (sodium dodecyl sulfate) and 0.8 ml of Triton X-100 (polyethylene glycol p-isooctylphenyl ether) to 200 ml of low-salt buffer.

[0061] Steps:

[0062] 1) Take (reindeer, eld, sambar, elk, fallow deer, white-lipped deer) 200 μl of anticoagulant blood and 400 μl of low-salt buffer solution into a 1.5 ml centrifuge tube, mix well, and centrifuge at 1 000 RPM at room temperature for 10 min, discard the supernatant;

[0063] 2) Add 500 μl low-salt buffer, mix thoroughly, centrifuge at 1 000 RPM for 5 min at room temperature, and repeat the previous step;

[0064] 3) Carefully aspirate ...

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Abstract

The invention discloses a method for safely and quickly extracting a genomic deoxyribose nucleic acid (DNA) from blood. The method comprises the following steps of: lysing a blood cell by using TritonX-100 and sodium dodecyl sulfonate (SDS), precipitating a protein with saturated NaCl, separating the protein through centrifugation, and reducing the mechanical shearing damage of a solution to the DNA; preventing and inhibiting the degradation of deoxyribonuclease (DNase) to the DNA; excluding the pollution caused by organic solvent, metal ions and other cnucleic acid molecules; and reducing the protein, polyoses, lipids and the like to the lowest degree, wherein the extracted DNA has certain length, and substances which inhibit enzymes exist in the genomic DNA after purification. The method for extracting the genomic DNA from the animal blood is harmless, quick and simple in operation.

Description

technical field : [0001] The invention discloses a method for safely and rapidly extracting genomic DNA from blood, relates to the preparation technology of blood total DNA, and belongs to the technical research field of animal molecular biology. Background technique: [0002] Genomic DNA (genomic DNA) is the carrier of genetic information, the most important biological information molecule, the main object of molecular biology research, the main material basis for genetic manipulation, and plays an important role in genomics research. Deoxyribose nucleic acid (Deoxyribonucleic Acid, DNA) is a kind of biological macromolecule with genetic information, which is composed of four main deoxynucleotides (dAMP, dGMP, dCMT and dTMP) linked by 3′,5′-phosphodiester bonds made. Their composition and arrangement are different, showing different biological functions, such as coding function, regulation function of replication and transcription, etc. For sequencing, hybridization and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 邢秀梅杨福合査代明荣敏苏伟林吴琼
Owner 邢秀梅
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