Huang C-W, Huang M-C
Department of Animal Science, National Chung Hsing University, Taichung, Taiwan.
Reprod Domest Anim. 2008 Dec;43(6):643-6. doi: 10.1111/j.1439-0531.2007.00938.x. Epub 2008 Oct 15.
The aim of the present study is to identify ostrich sex by using polymerase chain reaction (PCR) on micro amounts of DNA from blood, bloodstain and feathers. Sixteen male and 18 female ostriches were used as test individuals. Genomic DNA as a template was extracted by the Chelex method. Ostrsex-P1 and P2 primers were designed to perform PCR amplification on the template. PCR products were checked using agarose gel electrophoresis with ethidium bromide staining and ostrich sex was determined directly by the bands shown on the gel. The results demonstrate that ostrich sex can be determined by the extraction of DNA from as little as 0.0125 microl blood using Chelex, whereby the use of large amounts of organic solvents such as phenol and chloroform are unnecessary. In addition, it is possible to identify ostrich sex using micro amounts of DNA extracted from bloodstains and/or feathers. The use of feathers particularly avoids unwanted sampling problems such as the difficulty of collecting ostrich blood, the stress to the ostrich caused by bleeding, and the demand for a lot of manpower for ostrich restraint.
本研究的目的是通过对微量血液、血迹和羽毛中的DNA进行聚合酶链反应(PCR)来鉴定鸵鸟的性别。使用16只雄性鸵鸟和18只雌性鸵鸟作为测试个体。采用Chelex法提取基因组DNA作为模板。设计Ostrsex-P1和P2引物对模板进行PCR扩增。使用溴化乙锭染色的琼脂糖凝胶电泳检查PCR产物,并直接根据凝胶上显示的条带确定鸵鸟的性别。结果表明,使用Chelex从低至0.0125微升血液中提取DNA即可确定鸵鸟性别,因此无需使用大量有机溶剂如苯酚和氯仿。此外,使用从血迹和/或羽毛中提取的微量DNA也能够鉴定鸵鸟性别。使用羽毛尤其避免了不必要的采样问题,如采集鸵鸟血液困难、采血对鸵鸟造成的应激以及需要大量人力来约束鸵鸟等。