Abstract
New DNA sequencing technologies are allowing researchers to explore the genomes of the millions of natural history specimens collected prior to the molecular era. Yet, we know little about how well specific next-generation sequencing (NGS) techniques work with the degraded DNA typically extracted from museum specimens. Here, we use one type of NGS approach, sequence capture of ultraconserved elements (UCEs), to collect data from bird museum specimens as old as 120years. We targeted 5060 UCE loci in 27 western scrub-jays (Aphelocoma californica) representing three evolutionary lineages that could be species, and we collected an average of 3749 UCE loci containing 4460 single nucleotide polymorphisms (SNPs). Despite older specimens producing fewer and shorter loci in general, we collected thousands of markers from even the oldest specimens. More sequencing reads per individual helped to boost the number of UCE loci we recovered from older specimens, but more sequencing was not as successful at increasing the length of loci. We detected contamination in some samples and determined that contamination was more prevalent in older samples that were subject to less sequencing. For the phylogeny generated from concatenated UCE loci, contamination led to incorrect placement of some individuals. In contrast, a species tree constructed from SNPs called within UCE loci correctly placed individuals into three monophyletic groups, perhaps because of the stricter analytical procedures used for SNP calling. This study and other recent studies on the genomics of museum specimens have profound implications for natural history collections, where millions of older specimens should now be considered genomic resources.
Highlights
Natural history collections house billions of specimens worldwide, providing a record of Earth’s biodiversity in space and time and a source of data to answer questions about evolution, ecology, conservation, and human health (Austin &Melville, 2006; Suarez & Tsutsui, 2004; Winker, 2004)
DNA reacted differently than DNA from fresh samples when varying certain parameters of the sequence capture protocol. They found that decreasing the hybridization temperature had a positive effect on capture efficiency for DNA from fresh tissue, but a negative effect on ancient DNA
This study confirms that ancient DNA might not always react as we expect with protocols developed for DNA extracted from fresh samples
Summary
Natural history collections house billions of specimens worldwide, providing a record of Earth’s biodiversity in space and time and a source of data to answer questions about evolution, ecology, conservation, and human health (Austin &Melville, 2006; Suarez & Tsutsui, 2004; Winker, 2004). Modern specimen preparation methods include archiving frozen tissue for DNA analysis, yet the millions of specimens collected prior to the molecular age lack this ready source of high quality DNA. Next-generation DNA sequencing (NGS) offers a more efficient way of sequencing DNA from museum specimens (Hofreiter et al, 2015; Rizzi et al., 2012). First applied to sequence the mammoth genome (Poinar et al, 2006), scientists have used NGS to sequence the Neanderthal genome from bone fragments, taking advantage of the millions of sequence reads to overcome the problem of contamination from modern humans that would otherwise have reduced the overall number of Neanderthal DNA reads to an unacceptably low number (Green et al, 2010; Meyer et al, 2012)
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