Multiplex polymerase chain reaction (PCR) primers and method for identifying genotypes of green-shell laying hens

Abstract

The invention discloses multiplex polymerase chain reaction (PCR) primers and a method for identifying the genotypes of green-shell laying hens. Three primers F, R1 and R2 are firstly designed, wherein F and R2 are fragments in a SLCO1B3 gene of a chicken normal genome. If an individual does not carry an insert fragment, the F and the R2 are paired, a single purpose band of 397 bp is only amplified, and no green-shell egg is produced. The R1 is a fragment of an EAV-HP insertion sequence; if the individual carries the insert fragment, the F and the R1 are paired, a single purpose band of 223 bpis only amplified, and green-shell eggs are produced. If one chromosome of the individual carries the insert fragment and the other one chromosome does not carry the insert fragment, two purpose bands of 223 bp and 397 bp are amplified, and green-shell eggs are still produced. Compared with other identification methods, the primer combination enables the two bands in heterozygote to be more clearly separated, and is beneficial to increment of the genotype identification accuracy of the individual to be tested.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a multiplex PCR primer and an identification method for identifying the genotype of green-shell laying hens. Background technique [0002] Egg shell color is an important egg trait and economic trait. The depth of egg shell color directly affects the sales of eggs, thereby affecting the economic benefits of farmers. Green-eggs are widely welcomed because of their color in line with the current needs of people in pursuit of health. In my country, only a few local chicken breeds such as Dongxiang green-shell hens and Changshun green-shell hens produce green-shell eggs, which have become valuable genetic resources. A number of domestic breeding companies have also introduced green-shell egg genes to carry out crossbreeding and cultivate specialized strains of green-shell layer hens for commercial production. However, in actual production, since heterozygous hens can also lay green egg...

AI Technical Summary

Problems solved by technology

[0004] Although the currently reported patented method of using molecular markers to identify the genotype of green-shelled eggs can identify the genotype of green-shelled eggs, some methods require secondary amplification, and some methods amplify the two target fragments in the agarose gel. There are problems such as that the bands are relatively close to each other, resulting in unclear separation of some individuals

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