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Method for extracting whole blood DNA of bird species

A technology for poultry and blood, applied in the field of DNA extraction of whole blood of poultry, can solve the problems of long extraction time, non-renewable, limited sources, etc., and achieve the effect of short extraction time

Inactive Publication Date: 2010-06-23
SICHUAN AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0020] 2. The extraction time is longer:
[0023] In the actual operation process, blood sample resources are very precious, and the source is limited and non-renewable. Losing it will cause inestimable losses, and the consequences are irreversible.

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  • Method for extracting whole blood DNA of bird species
  • Method for extracting whole blood DNA of bird species
  • Method for extracting whole blood DNA of bird species

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Embodiment Construction

[0042] The specific steps of the present invention are as follows: before the test, the used utensils, reagents and consumables are autoclaved. Specific steps are as follows:

[0043] ① After the frozen blood is thawed at room temperature, take 22 μl of blood and add it to a centrifuge tube, then add 860 μl of buffer + 100 μl of lysate + 15 μl of proteinase K, the total volume does not exceed 1000 μl; vortex for 5 minutes to make it completely mixed;

[0044] ② 56 ℃ metal bath (water bath) digestion for more than 3 hours;

[0045] ③ Add 500 μl of pre-cooled (-20°C) chloroform, swirl and mix for 5 minutes to make it completely mixed;

[0046] ④Centrifuge at 13000r / min for 5 minutes, transfer the upper aqueous phase (500-600μl) to another clean centrifuge tube, be careful not to stir or absorb the lower two phases;

[0047] ⑤ Add an equal volume of pre-cooled (-20°C) isopropanol to the supernatant, and shake the centrifuge tube slowly for 5 minutes;

[0048] ⑥Centrifuge at 13...

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Abstract

The invention relates to a method for extracting the whole blood DNA of bird species, which comprises the following steps: taking 22 mu l of blood to add into a centrifugal tube after the frozen blood is melted at the room temperature; and then, adding 860 mul of buffer solution, 100 mul of schizolysis solution and 15 mu l of prolease K with the total volume not exceeding 1000 mul. After the DNA is obtained by the extraction method of the invention, DNA sediments are dried by being placed in a vacuum drier or a baking oven with the temperature of 45 DEG C, the DNA is fully dissolved through adding 100 to 200 mu l TE solution according to the size of the sediments after a block mass becomes transparent, and the obtained materials are stored in a refrigerator at the temperature of 20 DEG C below zero. The extraction method does not relate to the use of virulent drugs of phenol and the like in the whole extraction process, and has no injury on bodies of operators. Compared with the existing extraction method, the invention has the advantages that the extraction method is safer, the minimum time consumption of the completion of the whole extraction process is within four hours, the extraction time is short, and the extraction is faster than that of the existing extraction method. The DNA extraction can be completed only through a micro amount of anticoagulation by using the extraction method of the invention, and the method is more efficient than the existing method.

Description

technical field [0001] The invention relates to a method for extracting poultry whole blood DNA. Background technique [0002] Genomic DNA contains all the genetic information in cells. In molecular experiments such as PCR detection and HLA genotyping, the quality of template genomic DNA preparation is the key to the success of the experiment. There are many methods for extracting genomic DNA from poultry blood. It is an important condition for extracting genomic DNA to choose a method that is simple, fast, low in price, good in purity and high in yield. However, the DNA extraction methods currently used take a long time to extract, and the DNA extraction method commonly used in molecular laboratories is the "phenol-chloroform" method. [0003] The specific steps of the "phenol-chloroform" method are as follows: [0004] Before the test, the utensils, reagents, and consumables used were autoclaved. [0005] ① After the frozen blood is thawed at room temperature, take 50 μ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C07H21/04C07H1/08
Inventor 周兵尹华东朱庆刘益平
Owner SICHUAN AGRI UNIV
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