A Long PCR–Based Approach for DNA Enrichment Prior to Next-Generation Sequencing for Systematic Studies

Abstract

Premise of the study: 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. 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.

Publisher

Botanical Society of America

Publication Date

1-7-2014

Publication Title

Applications in Plant Sciences

Department

Biology

Document Type

Article

DOI

https://dx.doi.org/10.3732/apps.1300063

Keywords

Angiosperms, Chloroplast enrichment, Long PCR, Next-generation sequencing, Plastome, Universal chloroplast PCR primers

Language

English

Format

text

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