Method for distinguishing green shell egg laying chicken and chicken green shell egg genotype as well as special fragment and primers

Abstract

The invention discloses a method for distinguishing green shell egg laying chicken and chicken green shell egg genotype as well as a special fragment and primers. The invention provides a method for identification or auxiliary identification of whether chicken to be detected is green shell egg chicken or not, in order to detect the genome DNA (deoxyribonucleic acid), if objective DNA molecules (sequence 4) are contained in the genome DNA, the chicken to be detected belongs to or is candidated as the green shell egg chicken, and when the objective DNA molecules are not contained in the genome DNA, the chicken to be detected does not belong to or is not candidated as the green shell egg chicken. Experiments prove that a 425bp short fragment is adopted, three primers are designed and synthesized according to the short fragment and are used for multiple PCR (polymerase chain reaction) amplification, whether the chicken to be detected lays green shell eggs or not can be distinguished through detecting whether amplification products contain the short fragment or not, and meanwhile, the chicken green shell egg genotype can be distinguished.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for identifying the genotype of green-shell laying hens and green-shell eggs, special fragments and primers. Background technique [0002] Eggshell color is one of the factors that affect egg quality. In the poultry egg market, eggs with smooth surface, beautiful color and uniformity are often favored by consumers. Green-shell eggs are popular in the poultry egg market for their unique eggshell color, and often appear in the high-end poultry egg market as "healthy eggs" and "healthy eggs". The traditional breeding of green-shell hens has the problems of impure selection and large variation in eggshell color. [0003] The green-shelled egg trait of chicken is a quality trait controlled by a dominant single gene (O), and both dominant homozygous (O / O) and heterozygous (O / o) individuals lay green-shelled eggs. This genetic characteristic determines that it is difficult to di...

AI Technical Summary

Problems solved by technology

This method can accurately distinguish green-shell laying hens from non-green-shell laying hens, and can determine whether the genotype of green-shell laying hens is dominant homozygous (O / O) or heterozygous (O / o), However, this method has three disadvantages: (1) Genotype detection takes a long time, because the length of the PCR amplified fragment is nearly 6kb including the sequences on both sides of the insert fragment, and the reaction time of one PCR reaction needs at least 4.5h; (2) Amplification The failure rate is high, and long-fragment PCR requires high template quality and amplification conditions, and a little carelessness can lead to reaction failure; (3) high cost, long-fragment PCR requires a dedicated long-fragment Taq enzyme, which is more expensive than ordinary Taq enzyme is 4-5 times higher

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