Data from: A long PCR based approach for DNA enrichment prior to next-generation sequencing for systematic studies

Premise of the study: We present an alternative approach for molecular systematic studies that combines long PCR and next-generation sequencing (NGS). Our approach can be used to generate templates from any DNA source for NGS. Here we test our approach by amplifying complete chloroplast genomes and we present a set of 58 potentially universal primers for angiosperms to do so. Additionally, this approach is likely to be particularly useful for nuclear regions. Methods and Results: Chloroplast genomes of 30 species across angiosperms were amplified to test our approach. Amplification success varied depending on whether PCR conditions were optimized for a given taxon. To further test our approach, some amplicons were sequenced on an Illumina HiSeq 2000. Conclusions: Although here we tested this approach by sequencing plastomes, long PCR amplicons could be generated using DNA from any genome, expanding the possibilities of this approach for molecular systematic studies.

研究背景：我们提出了一种结合长聚合酶链式反应（long PCR）与下一代测序（next-generation sequencing，NGS）的分子系统学研究替代方案。该方案可从任意DNA来源中为NGS实验制备测序模板。本研究通过扩增完整叶绿体基因组对该方案进行验证，并提供了一套适用于被子植物（angiosperms）的58条通用引物。此外，该方案对核基因区域的研究亦具备显著应用潜力。方法与结果：本研究扩增了覆盖被子植物（angiosperms）的30个物种的叶绿体基因组以验证方案可行性。扩增成功率因类群是否适配优化后的PCR反应条件存在差异。为进一步验证方案，部分扩增产物通过Illumina HiSeq 2000平台完成测序。结论：尽管本研究通过测序质体基因组（plastomes）验证了该方案，但长PCR扩增产物可从任意基因组的DNA中制备，进一步拓展了该方案在分子系统学研究中的应用范围。