This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Amplification of Ultra-Trace DNA from Early Sheep Embryos Based on qPCR: Establishing a Gender Identification System
by
, ,
Peng Niu
1
,
Weikun Tao
2,
Fei Huang
1,
Xiaopeng Li
1
,
Xueyan Wang
1,
Jie Wang
1,3,
Qinghua Gao
1,3 and
Di Fang
1,3,* 1
College of Life Science and Technology, Tarim University, Alar 843300, China
2
College of Animal Science and Technology, Shihezi University, Shihezi 832003, China
3
Key Laboratory of Tarim Animal Husbandry Science and Technology, Xinjiang Production &Construction Corps; Alar 843300, China
*
Author to whom correspondence should be addressed.
Biology 2025, 14(9), 1144; https://doi.org/10.3390/biology14091144
Submission received: 25 June 2025
/
Revised: 19 August 2025
/
Accepted: 25 August 2025
/
Published: 29 August 2025
(This article belongs to the Special Issue The Biology of Animal Reproduction)
Simple Summary
Identifying the sex of early mammalian embryos is key for gender control. This study used ultra-trace amounts of sheep embryonic DNA templates and specific probes to amplify the SRY gene, establishing a time-saving, effective, simple, and reliable embryo sex identification method. The results showed that the accuracy of embryo sex identification was 100%. Combined with embryo transfer, this technology can accelerate breed selection and promote the development of embryo commercialization.
Abstract
The ability to identify and control the sex of embryos holds significant commercial implications for livestock production. Due to the high costs and instability of detection methods, it is necessary to establish a time-saving, effective, simple, and reliable method to identify the sex of mammalian embryos. This study designs primer probes based on the SRY gene sequence in the sex-determining region of the Y chromosome of sheep and combines qPCR to amplify both the SRY gene and the internal reference gene GAPDH as a control. By adjusting and optimizing the detection system with ultra-trace of genomic DNA (gDNA) from sheep blood and embryos, we sexed sheep embryos cultured in vitro. The results show that male embryos exhibit specific amplification bands for both the GAPDH and SRY genes, while female embryos only have amplification bands for the GAPDH gene. The sensitivity of the sheep embryo sex identification system established in this study can be as low as 5.64 pg of DNA, with an accuracy rate of 100% for embryo sex identification.
