Subba Prakrit, Adeniran-Obey Saidat O, Kraft Fanny-Linn, Chapman Susan C, Shay Natalie A, Liedl Shannon R, Wild Mark D, Wolcott Scarlett A, George Julia M
Department of Biological Sciences and Center for Human Genetics, Clemson University, Clemson, SC 29634, USA.
Centre for Integrative Ecology, School of Life and Environmental Sciences, Deakin University, Geelong, 3228 Victoria, Australia.
bioRxiv. 2025 Aug 28:2025.08.27.671803. doi: 10.1101/2025.08.27.671803.
Studies of early development in birds typically rely on PCR analysis of genomic DNA to identify embryonic or neonatal sex. In zebra finches and other birds, males are the homogametic sex (ZZ) while females are heterogametic (ZW), and females are distinguished by the presence of specific sequences on the female-specific W chromosome. However, when only a single W locus is analyzed, lack of a PCR product in a sample could potentially arise from genetic variation or technical failure of the amplification, leading to false identification of female samples as males. To mitigate this concern, we developed an approach that targets two different W loci, using SYBR-based quantitative PCR to analyze amplification curves. We applied this method to determine sex of 30 zebra finch embryos (embryonic day 13) and subsequently confirmed genetic sex by brain transcriptome sequencing. We also identified and tested primer sets that are effective for sex determination in chickens.
对鸟类早期发育的研究通常依靠对基因组DNA进行PCR分析来鉴定胚胎或幼鸟的性别。在斑胸草雀和其他鸟类中，雄性是同配性别（ZZ），而雌性是异配性别（ZW），雌性可通过雌性特异性W染色体上特定序列的存在来区分。然而，当只分析一个W基因座时，样本中PCR产物的缺失可能是由于基因变异或扩增技术失败导致的，从而导致将雌性样本错误鉴定为雄性。为了减轻这一担忧，我们开发了一种针对两个不同W基因座的方法，使用基于SYBR的定量PCR来分析扩增曲线。我们应用这种方法来确定30只斑胸草雀胚胎（胚胎发育第13天）的性别，随后通过脑转录组测序确认了遗传性别。我们还鉴定并测试了对鸡性别鉴定有效的引物组。
