Kit and method for rapidly extracting blood DNA in batches by adopting saturated sodium chloride process

A kit and sodium chloride technology, applied in the field of molecular biology, can solve the problems of unfavorable health of experimenters, affecting the speed of analysis and detection, low DNA yield, etc., to achieve easy purchase, convenient operation, and high DNA content. Effect

Inactive Publication Date: 2015-04-29
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the spin column method can extract DNA and ensure its purity, the yield of DNA extracted by it is generally not high, which cannot meet the requirements of large-scale animal molecular genetic analysis, forensic testing, archaeological analysis, and medical testing.
In addition, the spin column method uses some toxic reagents, such as phenol, chloroform, etc., which is not conducive to the health of the experimenters; at the...

Method used

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  • Kit and method for rapidly extracting blood DNA in batches by adopting saturated sodium chloride process
  • Kit and method for rapidly extracting blood DNA in batches by adopting saturated sodium chloride process
  • Kit and method for rapidly extracting blood DNA in batches by adopting saturated sodium chloride process

Examples

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Embodiment 1

[0033] The kit for rapid extraction of genomic DNA from blood described in this invention patent contains erythrocyte lysate, deproteinized solution, protein precipitation solution, adjusting binding solution, eluent and 20mg / ml proteinase K solution.

[0034] RBC lysate contains NH 4 Cl, KHCO 3 and Na 2 EDTA, pH 7.2-7.4; specific preparation method: NH 4 Cl 8.29g, KHCO 3 31g. Na 2 37.2 mg of EDTA was dissolved in 1000 mL of distilled water, added HCl to adjust the pH to 7.2-7.4, and sterilized under high temperature and high pressure for use.

[0035] The protein-removing solution is 10% SDS (10% sodium dodecylsulfonate); the specific preparation method: weigh 2g of SDS, add 20ml of ultrapure water, heat to 68°C to aid dissolution, add a few drops of HCl to adjust the pH to 7.2, High temperature and high pressure sterilization for use.

[0036] The protein-removing precipitation solution is saturated NaCl; the specific preparation method: add NaCl to 100ml of water until...

Embodiment 2

[0040] Example 2: Rapid extraction of genomic DNA from chicken blood

[0041] Genomic DNA was rapidly extracted from chicken blood using the kit described in Example 1.

[0042] 1. Extraction reagent

[0043] Red blood cell lysate: contains NH 4 Cl, KHCO 3 and Na 2 EDTA, pH 7.2-7.4;

[0044] Protein removal solution: 10% SDS;

[0045] Protein precipitation solution: saturated NaCl;

[0046] Adjust binding solution: isopropanol; eluent: 70% ethanol;

[0047] Proteinase K: Proteinase K solution at a concentration of 20 mg / ml.

[0048] 2. Extraction method

[0049] (1) Use a pipette gun to draw 10ul chicken blood sample and place it in a 1.5ml EP tube;

[0050] (2) Add 400ul erythrocyte lysate, 20ul proteinase K (20mg / ml), 25ul deproteinized solution 10% SDS, vortex and mix well.

[0051] (3) Put the EP tube into a water bath at about 56°C for 3 to 6 hours (increase the digestion time or even overnight digestion depending on the situation, try to digest the sample compl...

Embodiment 3

[0056] Embodiment 3: Genomic DNA is extracted rapidly from the blood of forest musk deer

[0057] The kit described in Example 1 was used to rapidly extract genomic DNA from the blood of musk deer.

[0058] 1. Extraction reagent

[0059] Red blood cell lysate: contains NH 4 Cl, KHCO 3 and Na 2 EDTA, pH 7.2-7.4;

[0060] Protein removal solution: 10% SDS;

[0061] Protein precipitation solution: saturated NaCl;

[0062] Adjust binding solution: isopropanol; eluent: 70% ethanol;

[0063] Proteinase K: Proteinase K solution at a concentration of 20 mg / ml.

[0064] 2. Extraction method

[0065] (1) Use a pipette gun to draw 10ul forest musk deer blood sample and place it in a 1.5ml EP tube;

[0066] (2) Add 400ul erythrocyte lysate, 20ul proteinase K (20mg / ml), 25ul deproteinized solution 10% SDS, vortex and mix well.

[0067] (3) Put the EP tube into a water bath at about 56°C for 3 to 6 hours (increase the digestion time or even overnight digestion depending on the sit...

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Abstract

The invention provides a kit and a method for rapidly extracting blood DNA in batches by adopting a saturated sodium chloride process. The kit contains red blood cell lysate, a deproteinization solution, a protein precipitation solution, a regulation binding solution, an eluant and a proteinase k solution, wherein the red blood cell lysate contains 8.29g/L NH4Cl, 31g/L KHCO3 and 0.37g/L Na2EDTA, and pH value is 7.2-7.4; the deproteinization solution is 20% SDS; the protein precipitation solution is saturated NaCl; the regulation binding solution is isopropyl alcohol; the eluant is 70% ethyl alcohol; and concentration of the proteinase k solution is 20mg/ml. The method for extracting the blood DNA by utilizing the kit has the advantages that content of the extracted DNA is high, purity is high, extraction is rapid, no toxic reagent is adopted, PCR amplification detection is carried out on the extracted genome DNA, and the extracted blood DNA can meet the requirements of many follow-up molecular biology experiments.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a kit for rapidly extracting blood DNA by a saturated sodium chloride method, and also relates to a method for using the kit to extract DNA from blood samples. Background technique [0002] The genomic DNA extracted from different sources of material is very different due to the difference in the starting amount of the material, the cell structure and its components. Therefore, sufficient material input is critical to extract enough genomic DNA for subsequent experiments. In many zoological experiments, a large amount of genomic DNA can be obtained by collecting a small amount of animal blood, and the collection method is fast, so that a lot of molecular genetic breeding research can be carried out. Therefore, the effective extraction of genomic DNA from blood samples becomes a key step to achieve the goals in the above fields. [0003] At present, there are various methods for...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 李地艳张璞杨明耀徐怀亮朱庆陈彬龙胡耀东吴楠张龙施贤
Owner SICHUAN AGRI UNIV
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