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Y 染色体捕获方法学:使用流式细胞术、激光显微切割和磁珠链霉亲和素对 Y 染色体进行全基因组测序。

Methodology for Y Chromosome Capture: A complete genome sequence of  Y chromosome using flow cytometry, laser microdissection and magnetic streptavidin-beads.

机构信息

GENYO (Pfizer-University of Granada-Andalusian Government Centre for Genomics and Oncological Research), Av. Ilustracion, 114 - PTS - 18016, Granada, Spain.

Centro di Antropologia Molecolare per lo Studio del DNA Antico, Dipartimento di Biologia, Università degli Studi di Roma"Tor Vergata", via della Ricerca Scientifica n. 1, 00173, Rome, Italy.

出版信息

Sci Rep. 2018 Jun 21;8(1):9436. doi: 10.1038/s41598-018-27819-x.

Abstract

This study is a comparison of the efficiency of three technologies used for Y chromosome capture and the next-generation sequencing (NGS) technologies applied for determining its whole sequence. Our main findings disclose that streptavidin-biotin magnetic particle-based capture methodology offers better and a deeper sequence coverage for Y chromosome capture, compared to chromosome sorting and microdissection procedures. Moreover, this methodology is less time consuming and the most selective for capturing only Y chromosomal material, in contrast with other methodologies that result in considerable background material from other, non-targeted chromosomes. NGS results compared between two platforms, NextSeq 500 and SOLID 5500xl, produce the same coverage results. This is the first study to explore a methodological comparison of Y chromosome capture and genetic analysis. Our results indicate an improved strategy for Y chromosome research with applications in several scientific fields where this chromosome plays an important role, such as forensics, medical sciences, molecular anthropology and cancer sciences.

摘要

本研究比较了三种用于 Y 染色体捕获的技术与应用于确定其全序列的下一代测序(NGS)技术的效率。我们的主要发现表明,与染色体分选和显微切割程序相比,基于链霉亲和素-生物素磁珠的捕获方法为 Y 染色体捕获提供了更好和更深的序列覆盖度。此外,与其他方法相比,该方法耗时更短,并且更具选择性,只能捕获 Y 染色体物质,而其他方法则会从其他非目标染色体产生大量背景物质。在 NextSeq 500 和 SOLID 5500xl 两个平台之间进行的 NGS 结果比较产生了相同的覆盖结果。这是首次探索 Y 染色体捕获和遗传分析的方法比较的研究。我们的结果表明,对于 Y 染色体研究的一种改进策略,该策略在法医、医学科学、分子人类学和癌症科学等 Y 染色体发挥重要作用的多个科学领域都有应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da7e/6013464/1ca0ea7bd855/41598_2018_27819_Fig1_HTML.jpg

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