Method for extracting genome DNA of poultry

Abstract

The invention discloses a method for extracting genome DNA of poultry, applied to large-scale collection, separation and purification of genome DNA of poultry in the actual production. Based on the condition that the poultry blood corpuscle contains a karyon, the method for extracting genome DNA of poultry in the invention proposes a principal that the DNA genome can be extracted once the poultry blood is taken out, completely saving the steps of anticoagulation and blood preservation; the method also proposes that blood is taken from the plantar vein, realizing simple and efficient blood sample collection and less stimulation to the poultry, and furthest avoiding reduction of egg and meal yield because of blood taking. The blood sample not more than 20mul in each poultry is taken on spot and then directly put into a 1.5ml centrifugal tube and mixed with the 700mul of poultry blood lysate in the centrifugal tube, put into an ice-cube box and taken back to the laboratory for DNA extraction; the whole process is secure and reliable, and biological security danger caused by massive poultry blood transport is reduced. Aiming at the problem that protein pollution easily occurs in the poultry blood extraction process, the method of the invention proposes that the DNA is extracted according to the original phenol method, but the saturated phenol is treated for two times and centrifuged at four degrees, thus the protein pollution is extremely reduced, the purification of the extracted DNA is much improved, and the subsequent molecular detection is much facilitated.

Description

technical field [0001] The invention relates to a method for extracting poultry genome DNA. Background technique [0002] Molecular genetic marker, also known as DNA molecular marker, refers to a DNA sequence located at a certain position on the chromosome, which can be stably inherited and can be detected. It is derived from the sequence polymorphism of DNA, which satisfies the law of Mendelian inheritance and is transmitted stably along with the replication of DNA. According to the linkage law, it can be known that DNA fragments with relatively close distances on chromosomes can be transmitted to the next generation of individuals in a chain, so DNA molecular markers are often used to track and determine the transmission law of a specific gene in an individual, and become a special manifestation of the genotype that is easy to identify form. Compared with cytogenetic markers, biochemical genetic markers and morphological genetic markers, molecular genetic markers are not...

AI Technical Summary

Problems solved by technology

In the on-site blood collection of chickens, a sterile syringe is generally used to collect blood from the vein under the wings of each chicken. This not only stresses the chickens, but also causes subcutaneous congestion, affects the health status of poultry individuals and even significantly reduces poultry egg production. Meat production, resulting in considerable economic and benefit losses; and because of the use of sterile syringes, the cost of blood collection under the wing is very high

Since it is necessary to accurately insert the needle into the subwing vein and draw blood, the operator has a high degree of proficiency, which undoubtedly increases the difficulty of blood drawing and the painful stimulation suffered by individual poultry, and the blood drawn must be checked on the spot. Anticoagulation treatment is also a time-consuming and labor-intensive project in the preliminary preparation

Due to the large amount of blood obtained from the subwing vein blood collection, generally up to 3-5ml, for poultry with nucleated blood cells, the average blood collection volume is much greater than the amount required for DNA extraction, which not only causes waste to a certain extent but also Increased adverse stimulation to individual poultry

And in practice, the anticoagulant cannot guarantee that each tube is in a purely anticoagulated state, which will cause uneven coagulation and cause differences in the amount of blood cells absorbed when extracting DNA, resulting in differences in the amount of DNA precipitated. Significantly, it brings inconvenience to the precise experiment of subsequent DNA

If hemolysis occurs in the anticoagulation process, there will be too few blood cells in the drawn blood sample, resulting in too little DNA precipitation or even an "empty tube" phenomenon that does not exist, resulting in rework, which not only affects the progress and effect of the work, but is even more serious Affects the next step in the laboratory DNA related testing work

To obtain genomic DNA from a large group of poultry, it is undoubtedly a time-consuming, labor-intensive, high-cost, and low-efficiency work to collect blood from the subwing vein and make anticoagulant blood in the laboratory.

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