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ddRAD 靶向富集测序在刺龙虾物种(短尾派蝦科:真龙虾属)中的效率。

Efficiency of ddRAD target enriched sequencing across spiny rock lobster species (Palinuridae: Jasus).

机构信息

Department of Ecology, Environment and Evolution, School of Life Sciences, La Trobe University, Melbourne, VIC 3086, Australia.

Institute for Marine and Antarctic Studies, University of Tasmania, Hobart, TAS 7001, Australia.

出版信息

Sci Rep. 2017 Jul 28;7(1):6781. doi: 10.1038/s41598-017-06582-5.

Abstract

Double digest restriction site-associated DNA sequencing (ddRADseq) and target capture sequencing methods are used to explore population and phylogenetic questions in non-model organisms. ddRADseq offers a simple and reliable protocol for population genomic studies, however it can result in a large amount of missing data due to allelic dropout. Target capture sequencing offers an opportunity to increase sequencing coverage with little missing data and consistent orthologous loci across samples, although this approach has generally been applied to conserved markers for deeper evolutionary questions. Here, we combine both methods to generate high quality sequencing data for population genomic studies of all marine lobster species from the genus Jasus. We designed probes based on ddRADseq libraries of two lobster species (Jasus edwardsii and Sagmariasus verreauxi) and evaluated the captured sequencing data in five other Jasus species. We validated 4,465 polymorphic loci amongst these species using a cost effective sequencing protocol, of which 1,730 were recovered from all species, and 4,026 were present in at least three species. The method was also successfully applied to DNA samples obtained from museum specimens. This data will be further used to assess spatial-temporal genetic variation in Jasus species found in the Southern Hemisphere.

摘要

双酶切限制位点相关 DNA 测序(ddRADseq)和靶向捕获测序方法被用于探索非模式生物的种群和系统发育问题。ddRADseq 为种群基因组学研究提供了一种简单可靠的方案,但由于等位基因缺失,它会导致大量数据缺失。靶向捕获测序为增加测序覆盖度提供了机会,同时在样本间具有很少的数据缺失和一致的同源基因座,尽管这种方法通常用于保守标记来解决更深层次的进化问题。在这里,我们结合了这两种方法,为研究属中的所有海洋龙虾物种的种群基因组学生成了高质量的测序数据。我们基于两种龙虾物种(爱德华氏真龙虾和罗氏沼虾)的 ddRADseq 文库设计了探针,并在另外五个 Jasus 物种中评估了捕获的测序数据。我们使用具有成本效益的测序方案在这些物种中验证了 4465 个多态性基因座,其中 1730 个存在于所有物种中,4026 个存在于至少三个物种中。该方法也成功应用于从博物馆标本中获得的 DNA 样本。这些数据将进一步用于评估南半球发现的 Jasus 物种的时空遗传变异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef3/5533801/c3cced3554d6/41598_2017_6582_Fig1_HTML.jpg

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