Cost-efficient high throughput capture of museum arthropod specimen DNA using PCR-generated baits

Gathering genetic data for rare species is one of the biggest remaining obstacles in modern phylogenetics, particularly for megadiverse groups such as arthropods. Next generation sequencing techniques allow for sequencing of short DNA fragments contained in preserved specimens >20 years old, but approaches such as whole genome sequencing are often too expensive for projects including many taxa. Several methods of reduced representation sequencing have been proposed that lower the cost of sequencing per specimen, but many remain costly because they involve synthesizing nucleotide probes and target hundreds of loci. These datasets are also frequently unique for each project and thus generally incompatible with other similar datasets.

Here, we explore utilization of in‐house generated DNA baits to capture commonly utilized mitochondrial and ribosomal DNA loci from insect museum specimens of various age and preservation types without the a priori need to know the sequence of the targe...

获取珍稀物种的遗传数据，是现代系统发育学（phylogenetics）尚存的最大障碍之一，对于节肢动物（arthropods）这类物种超多样化类群而言尤为如此。下一代测序（next generation sequencing）技术可对保存时长超过20年的标本所含的短DNA片段进行测序，但诸如全基因组测序（whole genome sequencing）这类方法，在涵盖众多分类单元（taxa）的研究项目中往往成本过高。已有多种简化基因组测序（reduced representation sequencing）方法被提出，用于降低单份标本的测序成本，但其中多数仍成本不菲——究其原因，这类方法需要合成核苷酸探针（nucleotide probes）并靶向数百个基因座（loci）。此外，这类数据集通常为单个研究项目所独有，因此普遍无法与其他同类数据集兼容。
本研究探讨利用实验室自制的DNA诱饵探针（DNA baits），从不同保存时长和保存方式的昆虫馆藏标本中捕获常用的线粒体DNA（mitochondrial DNA）与核糖体DNA（ribosomal DNA）基因座，且无需预先（a priori）知晓靶标序列（原文此处截断）。