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

染色体捕获方法:使用流式细胞术、激光显微切割和磁性链霉亲和素珠对 Y 染色体进行完整基因组序列分析

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作者:M J Alvarez-Cubero, O Santiago, C Martínez-Labarga, B Martínez-García, R Marrero-Díaz, A Rubio-Roldan, A M Pérez-Gutiérrez, P Carmona-Saez, J A Lorente, L J Martinez-Gonzalez

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.

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