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

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

Applications in Plant Sciences 2(1):

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. • 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. • 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.

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Accession: 051152607

Download citation: RISBibTeXText

PMID: 25202592

DOI: 10.3732/apps.1300063

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