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Whole genome DNA (Deoxyribonucleic Acid) extraction kit for blood and method thereof

A whole-genome and kit technology, which is used in blood whole-genome DNA extraction kits and its fields, to achieve the effects of improving accuracy, improving sensitivity, and reducing interference

Active Publication Date: 2014-09-03
ZICHENG RUISHENGHUI BEIJING BIOTECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The invention provides a kit for efficiently extracting whole-genome DNA from blood and a method for extracting whole-genome DNA from blood, so as to solve the problems of reducing the impact of organic solvents or strong alkalis on the health of operators and subsequent experiments, and the like. Efficient extraction of high-purity whole-genome DNA even from trace amounts of blood samples

Method used

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  • Whole genome DNA (Deoxyribonucleic Acid) extraction kit for blood and method thereof
  • Whole genome DNA (Deoxyribonucleic Acid) extraction kit for blood and method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0084] Whole Genome DNA Extraction Kit in Embodiment 1 Blood

[0085] The components of the kit are as follows:

[0086] 1) Red blood cell lysate: deionized ultrapure water (ddH 2 O), the resistivity is greater than 18MΩ*cm), sterilized at 121°C and 100 kPa pressure for 20 minutes; after sterilization, each 400ml is divided into 500ml sterile plastic bottles;

[0087] 2) White blood cell washing solution: add 50ml Triton-X100 to 400ml sterilized deionized ultrapure water, dissolve at 68°C, and set the volume to 500ml to prepare a 10% Triton-X100 mother solution; then add 10ml 10% Triton-X100 mother solution Add 990ml of sterilized deionized ultrapure water to prepare 0.1% Triton-X100 white blood cell washing solution; each 300ml is divided into 300ml sterile plastic bottles;

[0088] 3) Digestive solution: 60.55g Tris base dissolved in 400ml ddH 2 In O, use HCl to adjust the pH value to 8.0 (add about 2.0ml of concentrated HCl), and use ddH 2 O was fixed to 500ml, and ster...

Embodiment 2

[0101] Example 2 Using the kit of the present invention to extract whole genome DNA in frozen blood

[0102] 5ml EDTA anticoagulant blood was collected from 8 patients with clinical diabetic retinopathy and 7 normal people (a scientific research cooperation project with a cooperative unit, approved by the hospital ethics committee, and signed an informed consent form with the volunteers).

[0103] 1) Blood erythrocyte lysis: Take the frozen blood out of the -80°C refrigerator and put it in a 37°C water bath until the blood sample melts. Add 8ml of erythrocyte lysate and 1ml of blood sample to each 15ml centrifuge tube, shake and mix by hand, place at 4°C for 20min, then centrifuge at 2000rpm for 15min at low speed; after centrifugation, discard the supernatant;

[0104] 2) White blood cell washing: Add 2ml of white blood cell washing solution to the 15ml centrifuge tube containing the white blood cell pellet, mix well, put it into a low-speed horizontal centrifuge, centrifuge ...

Embodiment 3

[0111] Example 3 Using the kit of the present invention to extract whole genome DNA from bloodstains

[0112] Take 1ml of anticoagulant blood from 8 clinical patients and add them to worn clothes respectively, and place them at room temperature for 2-3 days. Cut off the blood-stained cloth, put them into 50ml centrifuge tubes with 30ml of erythrocyte lysate, vortex for 5 minutes, place at 4°C for 20 minutes, take out the cloth with tweezers, let the liquid drip dry as much as possible, and then centrifuge at 2000rpm at low speed 15min; discard the supernatant after centrifugation;

[0113] Add 30ml of leukocyte washing solution to the 50ml centrifuge tube containing leukocyte precipitate, mix well, centrifuge at low speed at 1000rpm for 10min, carefully discard the supernatant, and repeat washing twice; the rest of the steps follow step 3 in Example 2 -7 completed, the extracted whole genome DNA was detected by 0.8% agarose gel electrophoresis, figure 2 It shows that the wh...

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Abstract

The invention relates to a kit of extracting a whole genome DNA (Deoxyribonucleic Acid) from blood and a using method thereof. The kit is characterized by comprising a red blood cell lysate, a white blood cell scrubbing solution, digestive juice, proteinase K, a purifying liquid, gDNA salting out liquid, a gDNA scrubbing solution, a gDNA eluant and the like. The using method of the whole genome DNA extraction kit for blood is characterized by comprising the following steps: washing the red blood cell split to obtain the white blood cell; splitting the white blood cell by the digestive juice containing the proteinase K; and further purifying by an improved lithium chloride purifying liquid, salting out the liquid layer, and carrying out chromatography to obtain the high purity whole genome DNA. When the kit provided by the invention is used to extract the whole genome DNA in blood, plasma and serum in blood are not separated in advance but fresh or frozen anti-freezing whole blood is taken, wherein the lowest blood volume required reaches 20 microliters or blood cakes are required. According t the kit provided by the invention, the whole genome DNA with high purity can be fully unlinked and the PCR (Polymerase Chain Reaction) amplification is efficiently carried out, so that the kit is used for scientific research or clinical diagnostic analysis such as PCR amplification, gene expression, gene sequencing, whole genome sequencing, exome sequencing, gene mutation and single nucleotide polymorphism.

Description

technical field [0001] The invention relates to a kit for extracting whole genome DNA from blood and a method for extracting whole genome DNA from blood using the kit. When the kit of the present invention extracts the whole genome DNA in the blood, it is not necessary to separate the plasma and serum in the blood in advance, only need to take fresh or frozen anticoagulated whole blood, and the minimum required blood volume can reach 20 μl or blood spots . The high-purity whole genome DNA obtained by its efficient extraction can be used for scientific research or clinical diagnosis analysis such as PCR amplification, gene expression, gene sequencing, whole genome sequencing, exome sequencing, gene mutation and single nucleotide polymorphism . . Background technique [0002] Genomic DNA (gDNA) is all genetic information in haploid cells, including coding sequences and non-coding sequences. All life information can be found from whole genome DNA, including gene expression,...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 曾沃坦黄启明梁辰陈建勋郑英海
Owner ZICHENG RUISHENGHUI BEIJING BIOTECH DEV CO LTD
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