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用低精子输入的射出牛精子进行基因组和甲基化分析的简单工作流程。

Simple workflow for genome and methylation analyses of ejaculated bovine spermatozoa with low sperm input.

机构信息

Department of Animal Science, Michigan State University, East Lansing, MI 48879, USA.

Colorado Center for Reproductive Medicine, Lone Tree, CO 80124, USA.

出版信息

Biotechniques. 2020 Mar;68(3):155-158. doi: 10.2144/btn-2019-0121. Epub 2020 Jan 15.

Abstract

We developed a simplified workflow of gDNA extraction from ejaculated bovine sperm using a low total number of sperm and a short time frame that yields high-quality DNA suitable for downstream methylation and genome analyses. These techniques have broad implications in human biomedical sciences and agriculture, including clinical diagnoses of infertility, the identification of single-nucleotide polymorphisms and aberrant methylation patterns that can impact fertility, lower embryo development and contribute to heritable disease. The methods described here provide a reliable, simplistic approach for analyzing both the genomic and epigenomic status of whole sperm ejaculates that can be adapted for laboratory diagnostics, clinical reproductive practice and basic research.

摘要

我们开发了一种从射出的牛精子中提取 gDNA 的简化工作流程,该流程使用的精子总数较少,时间较短,但能获得高质量的 DNA,适用于下游甲基化和基因组分析。这些技术在人类生物医学科学和农业中有广泛的应用,包括不育症的临床诊断、单核苷酸多态性和异常甲基化模式的鉴定,这些模式可能会影响生育能力、降低胚胎发育,并导致遗传性疾病。这里描述的方法为分析整个精子射精液的基因组和表观基因组状态提供了一种可靠、简单的方法,可以适应实验室诊断、临床生殖实践和基础研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4231/7092705/1993ca1e74de/btn-68-155-g1.jpg

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