Development of SNP markers to identify the sex of the giant panda from naturally degraded fecal samples

Pub Date : 2023-11-21 DOI:10.1007/s12686-023-01326-2
Wei Xu, Daxing Xie, Jie Kou, Xiuyue Zhang, Yan Li, Ye Wang, Liang Zhang, Hong Liu, Jiawen Liu, Juan Wang, Li Luo, Xiaolan Wang, Kongju Wu, Fujun Shen
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Abstract

A non-invasive, fast, and reliable method for sex identification of the giant panda (Ailuropoda melanoleuca) was developed by genotyping five SNPs located at the sex chromosome amelogenin (AMEL) paralogs. The giant panda’s sex can be accurately distinguished by the homozygous or heterozygous SNP genotypes. Mass spectrometry assay and Sanger sequencing both successfully genotyped tissue and fresh fecal samples collected from known-sex giant pandas. Additionally, we simulated the conditions of wild fecal samples, which were naturally degraded for up to two weeks, and successfully genotyped these samples by mass spectrometry assay in parallel. These results suggest that this is a powerful method for sex determination in highly degraded and lower concentrated DNA samples.

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开发从自然降解的粪便样本中识别大熊猫性别的SNP标记
研究了一种无创、快速、可靠的大熊猫(Ailuropoda melanoleuca)性别鉴定方法,通过对性染色体amelgenin (AMEL)类群的5个snp进行基因分型。大熊猫的性别可以通过纯合或杂合SNP基因型准确区分。质谱分析和桑格测序都成功地对从已知性别的大熊猫收集的组织和新鲜粪便样本进行了基因分型。此外,我们模拟了自然降解的野生粪便样本的条件,并通过平行质谱分析成功地对这些样本进行了基因分型。这些结果表明,在高度降解和低浓度的DNA样品中,这是一种强有力的性别测定方法。
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