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| Construction of DAVID Technology for High-throughput and Low-cost Detection of Bovine (Bos taurus) MSTN Gene Mutations |
| PEI Dong-Chao, HAI Chao, ZHAO Chun-Sheng, ZHANG Li-Guo, WU Di, WANG De-Zheng, ZHANG Xin-Yi, YANG Lei*, LI Guang-Peng* |
| State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, Inner Mongolia University, Hohhot 010070, China |
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Abstract Myostatin (MSTN) is a negative regulator of skeletal muscle growth and development, and mutations in the MSTN gene can promote muscle development in animals. Currently, Sanger sequencing technology is the classic method for detecting MSTN gene mutations, but this method has the disadvantages of low detection throughput, low visualisation of mutation information and high detection cost. In this study, MSTN gene mutations in 96 cattle (Bos taurus) could be detected at one time by using next-generation sequencing technology with a specific Barcode sequence designed. In addition, based on the Barcode sequences in the sequencing Fastq result files, the related data analysis DAVID (detect and visualize insertion-deletion) software was developed using asynchronous threading and concurrent algorithms. Using DAVID software, the sequencing data of 96 mixed samples were split one by one and compared with amplicon sequence, and finally the graphical and statistical information of MSTN gene mutation was obtained. Using DAVID technology, 96 Mongolian cattle were examined, and found that 1 head of cattle was heterozygous for 11 bp deletion of the MSTN gene, and the remaining 95 head of cattle were wild-type, and the 11 bp heterozygous deletion was further verified by Sanger sequencing. This study constructed a high-throughput technical system for detecting bovine MSTN gene mutations, and provides technical support for genetic improvement and breeding of cattle.
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Received: 25 March 2024
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Corresponding Authors:
* mrknowall@126.com; gpengli@imu.edu.cn
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