【摘要】 根据绵羊SRY(sex determining region of the Y)基因的723 bp的高度保守序列设计跨度为193 bp的两对巢式雄性特异引物,同时根据绵羊β-B-珠蛋白基因设计跨度为278 bp的两对巢式引物为内标引物,通过析因设计法建立了巢式PCR体系,对公、母羊肝组织基因组进行巢式PCR扩增,经1.5%的琼脂糖电泳检测,公羊同时出现278 bp的β-B-珠蛋白基因扩增带和193 bp的雄性特异扩增带两条带,而母羊只扩增出278 bp常染色体基因序列扩增带.用此巢式PCR体系扩增淋巴细胞,灵敏度≥10 pg DNA(约3个细胞),鉴定全程不超过130 min.更多还原

【Abstract】 Two pairs of nested primers with the length of 193 bp were designed to identify the sex of early embryos based on the male specific SRY genes.Meanwhile,a pair of 278 bp primers were designed based on the sheep beta-B-globin genes on the autosome.The amplified product of the nested PCR by the 1.5 % agarose mini-gel electrophoresis showed that the male samples not only amplified a 193 base pair band but also 278 base pair band;the female samples amplified only a 278 base pair band.The developed nested PCR method was applied successfully with the sensitivity≥10 pg DNA(about 3 lymphocytes),and the identification processes was no longer than 130 minites.更多还原