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 nextgeneration sequencing. Our approach can be used to generate templates from any DNA source for next-generation sequencing. 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 and mitochondrial 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 PCK amplicons could be generated using DNA from any genome, expanding the possibilities of this approach for molecular systematic studies.