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首页> 美国卫生研究院文献>Journal of Equine Science >Whole-genome resequencing using genomic DNA extracted from horsehair rootsfor gene-doping control in horse sports
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Whole-genome resequencing using genomic DNA extracted from horsehair rootsfor gene-doping control in horse sports

机译：使用从Horsehair Roots提取的基因组DNA重新开始用于马运动中的基因掺杂控制

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Gene doping is prohibited in horseracing and equestrian sports. In previous studies, wedeveloped non-targeted transgene and genome editing detection methods based on wholegenome resequencing (WGR) using genomic DNA extracted from whole blood. In this study, weaimed to develop a WGR method using DNA extracts from hair roots. Hair roots are apreferred substrate because their collection is less invasive than blood collection. Hairis also easier to store for long periods of time. Although almost all genomic DNAextracted from hair root samples stored for years at room temperature was degraded, thequality of genomic DNA from samples stored for years at refrigerated temperatures (4–8°C)was maintained. High-molecular-weight genomic DNA was isolated from hair roots using amagnetic silica beads method of extraction, enabling WGR from horsehair root extracts.Nucleotide sequencing results and numbers of single-nucleotide polymorphisms andinsertions/deletions concurred with those previously reported for WGR of DNA extractedfrom whole blood. Therefore, we consider that storing hair samples at refrigeratedtemperatures prevents degradation of DNA, allowing the detection of gene doping in thesesamples based on WGR. It is likely this finding will also have a deterrent effect, as itis now possible to test horses with archived samples even if they or their parents aredeceased. To our knowledge, this is the first report employing WGR on horsehair rootsstored for a long term.
机译：猪蹄和马术运动中禁止基因掺杂。在以前的研究中，我们基于整体开发的非靶向转基因和基因组编辑检测方法基因组重新开始（WGR）使用从全血中提取的基因组DNA。在这项研究中，我们旨在使用来自毛根的DNA提取物进行WGR方法。头发根部是一个优选的基质，因为它们的收集比血液收集较少。头发长时间也更容易存储。虽然几乎所有基因组DNA从室温下储存多年的头发根样品中提取，降解了来自储存在冷藏温度（4-8°C）的样品的基因组DNA的质量被维持。使用a从毛根中分离高分子类别基因组DNA磁性二氧化硅珠萃取方法，从马毛根提取物中实现WGR。核苷酸测序结果和单核苷酸多态性的数量和数量与先前报告的DNA提取的DNA的插入/删除来自全血。因此，我们认为在冷藏时储存头发样品温度妨碍DNA的降解，从而检测这些基因掺杂基于WGR的样品。很可能这一发现也会有威慑效果，就像它一样现在可以用存档样本测试马匹，即使它们或父母也是如此死者。为了我们的知识，这是第一份雇用WGR在骑马根源的报告长期存放。

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