System, method and kit for identifying genders of cows and early embryos of cows

Abstract

The invention relates to a system, a method and a kit for identifying genders of cows and early embryos of the cows, and belongs to the field of biological technology detection. By utilizing the characteristic that bovine folded protein genes exist on X-chromosomes of the cows, a group of primers NB2-F and NB2-R and a probe NB2-P are designed. The group of probe is specially used for identifying the embryos of the cows, and other species cannot be identified, and then by utilizing the specific gene SRY of bulls, special design on the primers and the probe can be carried out by selecting sequence of the bulls, samples of the bulls only can be expanded by the designed primers BOV97M-F and BOV97M-R and the probe BOV97M-P, the two groups of probes are combined to build a composite detection system, so that the genders of the early embryos of the cows can be successfully identified. The system, the method and the kit for identifying the genders of the cows and the early embryos of the cowshave the advantages of high sensitivity (capable of detecting 3 to 5 cells), high sample processing speed, high detection speed, low cost and the like; in addition, pollution caused by multiple PCR (Polymerase Chain Reaction) and nested-PCR is effectively avoided, and a detection ratio and an accuracy ratio are up to 100 percent.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a sex identification method for cattle and early embryos thereof, which is an optimized upgrade and perfect supplement of the prior art. Background technique [0002] Certain production traits of animals are limited or affected by gender. For example, only cows in dairy cows show milk production traits, while male animals in beef cattle production have high economic value because of their fast growth. In order to give full play to the reproductive ability and The growth advantage of bulls and artificially controlling the sex of cattle have always been the goal that people pursue. Therefore, it is an important measure to improve the economic benefits of animal husbandry to quickly and accurately identify the sex of early embryos in livestock and implement sex control technology. [0003] In previous PCR sex identification, the housekeeping gene was used as the internal standard pr...

AI Technical Summary

Problems solved by technology

[0003] In previous PCR sex identification, the housekeeping gene was used as the internal standard primer, and the sex-specific SRY gene was used to design primers for double PCR amplification analysis. Template enrichment, cumbersome operation, time-consuming and labor-intensive, easy to pollute, technically difficult, not convenient for on-site operation

[0004] The single-plex PCR method is simple because it only amplifies the Y-chromosome-specific primers, and avoids the influence of the autosomal primers on the amplification of the Y-chromosome-specific primers in the multiplex PCR. Samples that do not react are misjudged as females, which increases the probability of false negative identification results. The traditional PCR method has a high demand for embryo samples. When the number of embryo cells is small, it is difficult to obtain templates after sample processing. Meet the requirements of PCR amplification for the template, that is, the nested PCR amplification is not good for the case of low template, mainly because the ordinary PCR method has certain requirements for the template concentration, the detection sensitivity is low, and electrophoresis detection is prone to contamination Therefore, developing a bovine embryo identification complex system with high detection sensitivity, high detection rate and high accuracy has become the current main task

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