miRNA marker related to pig hair-free character and application of miRNA marker
一种标志物、性状的技术,应用在miRNA标志物领域,能够解决耗时长、实验操作复杂等问题,达到结果准确、减小假阳性的效果
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Embodiment 1
[0040] Example 1: Screening for the gene miRNA-29a-5p associated with hairless traits
[0041] The experimental population of the present invention is a small amount of large white hairless pigs discovered earlier (discovery address: Anping County, Hengshui City, Hebei Province, discoverer: Ding Xiangdong, associate researcher of the Animal Science and Technology College of China Agricultural University)
[0042] S1. Hairless pig breeding plan: select normal boars and hairless sows to breed, and perform cesarean section when pregnant sows reach 41 days of gestation, take out pig embryos and collect two tissue pieces of size 2cm x 2cm on the back of the embryos, then separate the skin tissue and dry Disinfect. One of them was placed in liquid nitrogen for RNA preservation, and the other was placed in paraformaldehyde for later phenotypic identification.
[0043] S2, phenotype identification: phenotype identification adopts hematoxylin and eosin (HE) staining to carry out hair ...
Embodiment 2
[0054] Example 2 Verifying the relationship between miRNA-29a-5pRNA gene expression and hairless phenotype in pig embryo skin (experimental verification of the relationship between the value of miRNA-29a-5pRNA gene expression and hairless traits)
[0055] S1. Hairless pig breeding plan: select normal boars and hairless sows to breed, and perform cesarean section when pregnant sows reach 41 days of gestation, take out pig embryos and collect two tissue pieces of size 2cm x 2cm on the back of the embryos, then separate the skin tissue and dry Disinfect. Place samples in liquid nitrogen for RNA preservation.
[0056] S2, miRNA-29a-5p primer design:
[0057] The sequence information of the miRNA-29a-5p gene was obtained from the NCBI database (https: / / www.ncbi.nlm.nih.gov / ), and designed using the software miRprimer2. The primers for amplifying the housekeeping gene GAPDH were designed in the same way as above, and were synthesized by Shanghai Sangon Bioengineering Co., Ltd.
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