title: A long PCR-based approach for DNA enrichment prior to next-generation sequencing for systematic studies.
abstract: OBJECTIVE
  We present an alternative approach for molecular systematic studies that combines long PCR and next-generation 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. •
  
  
  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.
authors.affiliation: Department of Biological Sciences and Institute for Bioinformatics and Evolutionary Studies, University of Idaho, 875 Perimeter Drive MS 3051, Moscow, Idaho 83844-3051 USA ; College of Natural Resources, University of Idaho, 875 Perimeter Drive MS 1133, Moscow, Idaho 83844-1133 USA.
authors.forename: Simon
authors.initials: S
authors.lastname: Uribe-Convers
authors.affiliation: College of Natural Resources, University of Idaho, 875 Perimeter Drive MS 1133, Moscow, Idaho 83844-1133 USA.
authors.forename: Justin R
authors.initials: JR
authors.lastname: Duke
authors.affiliation: Department of Biology, Oberlin College, Science Center K111, 119 Woodland St., Oberlin, Ohio 44074-1097 USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Moore
authors.affiliation: Department of Biological Sciences and Institute for Bioinformatics and Evolutionary Studies, University of Idaho, 875 Perimeter Drive MS 3051, Moscow, Idaho 83844-3051 USA ; College of Natural Resources, University of Idaho, 875 Perimeter Drive MS 1133, Moscow, Idaho 83844-1133 USA.
authors.forename: David C
authors.initials: DC
authors.lastname: Tank
keywords: angiosperms; chloroplast enrichment; long PCR; next-generation sequencing; plastome; universal chloroplast PCR primers