Center for Conservation and Evolutionary Genetics, Smithsonian Conservation Biology Institute, National Zoological Park, Smithsonian Institution, PO Box 37012, Washington, DC 20013, USA.
Mol Ecol Resour. 2011 Sep;11(5):831-4. doi: 10.1111/j.1755-0998.2011.03030.x. Epub 2011 Jun 3.
We report the development of a reliable and efficient method for molecular sexing of all extant elephant taxa. We developed primers that amplify two short Y-specific fragments (SRY1 and AMELY2) and one longer X-specific fragment (PLP1), developed from elephant sequences in one multiplex PCR. All fragments were designed to be short (< 200 basepairs) for use with degraded DNA and to be 50 basepairs apart to optimize visualization on agarose gels or as electropherograms. The multiplex PCR method matched sexes for at least 97.9% of the noninvasive savannah elephant samples and produced the expected female/male banding patterns for 14 African forest and 11 Asian elephant samples. We found this method to be more robust, efficient and less prone to contamination than previously developed sexing methods for elephants.
我们报告了一种可靠且高效的分子性别鉴定方法，可用于所有现存的象类。我们开发了引物，可在一个多重 PCR 中扩增两个短的 Y 染色体特异性片段（SRY1 和 AMELY2）和一个较长的 X 染色体特异性片段（PLP1），这些片段均源自大象序列。所有片段的设计都较短（<200 个碱基对），以便用于降解的 DNA，并相距 50 个碱基对，以优化在琼脂糖凝胶或电泳图谱上的可视化。该多重 PCR 方法匹配了至少 97.9%的非侵入性萨凡纳象样本的性别，并产生了 14 头非洲森林象和 11 头亚洲象样本的预期雌性/雄性带型。我们发现，与以前开发的大象性别鉴定方法相比，这种方法更稳健、高效，且不易受到污染。
