Got Genome? A long PCR based method for chloroplast genome enrichment and phylogenomics in angiosperms

We present a straightforward and cost-effective long PCR based method to amplify whole chloroplast genomes from standard DNA extractions. Our method allows researchers to obtain large amounts of data from very small quantities of silica gel-dried or herbarium tissue (~0.02 g), as opposed to traditional plastome enrichment techniques that often require large quantities of fresh tissue (~15 g).

We designed 58 universal chloroplast primers that successfully amplify across the major lineages of angiosperms. From these, 16 primer combinations were chosen to amplify the complete chloroplast genome in overlapping fragments that ranged from 5-12 kb. For several genera in Orobanchaceae, we sequenced these amplicons on an Illumina HiSeq, but they could be sequenced using any traditional or next-generation sequencing technology.

Our method provides an affordable, straightforward and efficient method for amplifying complete chloroplast genomes across angiosperms from small quantities of tissue.

Note: This method was submitted to Application in Plant Sciences and includes a 'step-by-step' protocol, which can also be found at: http://www.simonuribe.com