Robust DNA isolation and high-throughput sequencing library construction for herbarium specimens

Herbaria are an invaluable source of plant material that can be used in a variety of biological studies. The use of herbarium specimens is associated with a number of challenges including sample preservation quality, degraded DNA, and destructive sampling of rare specimens. In order to more effectively use herbarium material in large sequencing projects, a dependable and scalable method of DNA isolation and library preparation is needed. This paper demonstrates a robust, beginning-to-end protocol for DNA isolation and high-throughput library construction from herbarium specimens that does not require modification for individual samples. This protocol is tailored for low quality dried plant material and takes advantage of existing methods by optimizing tissue grinding, modifying library size selection, and introducing an optional reamplification step for low yield libraries. Reamplification of low yield DNA libraries can rescue samples derived from irreplaceable and potentially valuable ...植物标本馆（Herbaria）是可用于各类生物学研究的珍贵植物材料来源。对植物标本馆样本的使用往往伴随诸多挑战，包括样本保存质量不佳、DNA降解以及对珍稀标本进行破坏性取样等。为了在大规模测序项目中更高效地利用植物标本馆材料，亟需一种可靠且可扩展的DNA提取与文库制备方法。本文展示了一套适用于植物标本馆样本的稳健全流程DNA提取及高通量测序文库构建方案，该方案无需针对单个样本进行调整。本方案专为低质量干燥植物材料优化，通过优化组织研磨步骤、调整文库片段大小筛选环节，并针对低产出文库引入可选重扩增步骤，以此充分利用现有技术方法。对低产出DNA文库进行重扩增，可拯救那些来自不可替代且极具潜在价值的……