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Rapid chick embryo egg sex identification method

An identification method and chicken embryo egg technology are applied in the fields of gender identification, developmental biology, tissue and cell engineering, and poultry breeding, which can solve problems such as difficulty in popularization, and achieve high identification accuracy, simple and repeatable sampling methods. strong effect

Inactive Publication Date: 2017-06-13
YANGZHOU UNIV
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  • Claims
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AI Technical Summary

Problems solved by technology

Although a large number of studies have shown that the PCR method is more feasible in the sex identification of chicken embryos, it still needs further processing after sample collection, and there are various problems that make it difficult to promote, and further optimization and improvement are needed

Method used

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  • Rapid chick embryo egg sex identification method
  • Rapid chick embryo egg sex identification method
  • Rapid chick embryo egg sex identification method

Examples

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Embodiment 1

[0029] (1) 16, 17, 18-day embryo egg allantoic fluid collection

[0030] Embryonic eggs hatch normally until the embryonic age of 16, 17, and 18 days. Use a 1ml syringe to collect allantoic fluid (sampling volume: 10 μL) through the tip of the embryonic egg (as shown in Figure 1), and wipe the sampling position of the embryonic egg with alcohol before sampling , sealed with paraffin after sampling (as attached figure 2 shown) to continue the incubation.

[0031] (2) CDH-W-specific primers for sex identification

[0032] Primers were designed according to the CDS sequence of CDH-W in the chicken NCBI database, and different fragments of ZW chromosome were amplified respectively. Males were double-banded (600bp and 450bp), and females were single-banded (450bp). The primer sequences are: Upstream Forward: 5'-TGCAGAAGCAATATTACAAG-3'; Downstream Reward: 5'-AATTCATTATCATCTGGTGG-3'. Using the obtained allantoic fluid as a template, PCR amplification was directly carried out, and ...

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Abstract

The invention discloses a novel rapid chick embryo egg sex identification method which comprises the following steps: hatching an egg to 16, 17 or 18 days of age, aseptically obtaining allantoic fluid (10 microliters) from the tip of the embryo egg through a 1 ml of injector, sampling and sealing with paraffin wax for normal hatching; taking the obtained allantoic fluid as a template, designing a CDH-W specific primer for PCR amplification, detecting the amplified primer through gel electrophoresis and performing sex identification, wherein double bands (600 bp and 450 bp) are adopted for male amplification, and a single band (450 bp) is adopted for female amplification; after normal hatching, statistically calculating the hatching rate and the death rate, and performing biostatistical analysis with a control group; comparing a sex identification result with an autopsy result of a hatched chick, performing correlation analysis, and statistically calculating the accuracy rate of sex identification (if reaching 99 percent, the accuracy rate is deemed to be 100 percent of effective).

Description

technical field [0001] The invention relates to the fields of poultry breeding, sex identification, developmental biology, tissue and cell engineering and the like, in particular to a method for rapid sex identification of chicken embryo eggs. Background technique [0002] Chicken sex identification is of great significance in the breeding industry and developmental biology research. In laying chickens, the economic value of hens is higher than that of males; in broiler chickens, the economic value of males is higher than that of females. chicken. At the same time, the domestic chicken is also a model animal for basic research and developmental biology of birds. Therefore, gender identification of chickens before hatching can greatly reduce costs, save manpower and material resources; it can also provide an experimental basis for related research. At present, there have been a large number of reports on the methods of sex identification of chicken embryos at home and abroa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2600/124C12Q2565/125
Inventor 李碧春靳锴左其生
Owner YANGZHOU UNIV
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