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一种基本的双酶ddRADseq方案在四个属的标本馆和硅胶干燥组织中表现良好。

A basic ddRADseq two-enzyme protocol performs well with herbarium and silica-dried tissues across four genera.

作者信息

Jordon-Thaden Ingrid E, Beck James B, Rushworth Catherine A, Windham Michael D, Diaz Nicolas, Cantley Jason T, Martine Christopher T, Rothfels Carl J

机构信息

University Herbaria and Department of Integrative Biology University of California Berkeley 3040 Valley Life Sciences Building Berkeley California 94720 USA.

Present address: Department of Botany University of Wisconsin 430 Lincoln Drive Madison Wisconsin 53706 USA.

出版信息

Appl Plant Sci. 2020 Apr 23;8(4):e11344. doi: 10.1002/aps3.11344. eCollection 2020 Apr.

Abstract

PREMISE

The ability to sequence genome-scale data from herbarium specimens would allow for the economical development of data sets with broad taxonomic and geographic sampling that would otherwise not be possible. Here, we evaluate the utility of a basic double-digest restriction site-associated DNA sequencing (ddRADseq) protocol using DNAs from four genera extracted from both silica-dried and herbarium tissue.

METHODS

DNAs from , , , and were processed with a ddRADseq protocol. The effects of DNA degradation, taxon, and specimen age were assessed.

RESULTS

Although taxon, preservation method, and specimen age affected data recovery, large phylogenetically informative data sets were obtained from the majority of samples.

DISCUSSION

These results suggest that herbarium samples can be incorporated into ddRADseq project designs, and that specimen age can be used as a rapid on-site guide for sample choice. The detailed protocol we provide will allow users to pursue herbarium-based ddRADseq projects that minimize the expenses associated with fieldwork and sample evaluation.

摘要

前提

对植物标本室标本进行基因组规模数据测序的能力,将使我们能够经济高效地开发具有广泛分类学和地理采样的数据集,否则这是不可能实现的。在此,我们使用从硅胶干燥和植物标本室组织中提取的来自四个属的DNA,评估了一种基本的双酶切限制性位点关联DNA测序(ddRADseq)方案的实用性。

方法

对来自[四个属的名称未给出]的DNA采用ddRADseq方案进行处理。评估了DNA降解、分类群和标本年代的影响。

结果

尽管分类群、保存方法和标本年代会影响数据回收,但大多数样本仍获得了大量具有系统发育信息的数据集。

讨论

这些结果表明,植物标本室样本可纳入ddRADseq项目设计中,并且标本年代可作为样本选择的快速现场指导。我们提供的详细方案将使用户能够开展基于植物标本室的ddRADseq项目,从而将与野外工作和样本评估相关的费用降至最低。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2459/7186894/627f9fdb5949/APS3-8-e11344-g001.jpg

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