2013
DOI: 10.2144/000114039
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Capturing Protein-Coding Genes Across Highly Divergent Species

Abstract: DNA hybridization capture combined with next generation sequencing can be used to determine the sequences of hundreds of target genes across hundreds of individuals in a single experiment. However, the approach has thus far only been successfully applied to capture targets that are highly similar in sequence to the bait molecules. Here we introduce modifications that extend the reach of the method to allow efficient capture of highly divergent homologous target sequences using a single set of baits. These modi… Show more

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Cited by 191 publications
(257 citation statements)
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References 29 publications
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“…In addition, a subset of the samples was also successfully subjected to a nuclear gene capture protocol (13). The inferred relationships among the four major clades based on a concatenated maximum-likelihood analysis of 100 single-copy nuclear genes was identical to the inference derived from the mitochondrial data (SI Appendix, Fig.…”
Section: Resultsmentioning
confidence: 78%
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“…In addition, a subset of the samples was also successfully subjected to a nuclear gene capture protocol (13). The inferred relationships among the four major clades based on a concatenated maximum-likelihood analysis of 100 single-copy nuclear genes was identical to the inference derived from the mitochondrial data (SI Appendix, Fig.…”
Section: Resultsmentioning
confidence: 78%
“…We prepared the target libraries following the methods of Li et al (13). For the subsequent target capture, we used "homemade baits," see SI Appendix, Detailed Protocol for Target Capture.…”
Section: Methodsmentioning
confidence: 99%
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“…Genomic DNA (0.5-3 ÎŒg per sample) was sheared to c. 500 bp using acoustic ultrasonication on a Covaris M220 Focused-ultrasonicator (Covaris, Inc., Massachusetts, USA) to form a target library for each specimen. Illumina sequencing libraries (Meyer & Kircher, 2010) were then prepared using the 'with-bead' method (Fisher et al, 2011), following Li et al (2013). Two custom biotinylated RNA bait libraries (MYbaits MYcroarray, Ann Arbor, MI, USA) were used in two separate DNA hybridization experiments per sample.…”
Section: Taxon Samplingmentioning
confidence: 99%
“…We sampled 19 species and 48 individuals ( Table 1, the DNA capture kit (MyBaits), which is able to support differences larger than 5% between 181 probe sequences and target sequences (e.g., Li et al, 2013). The complete sequence of the 182 chloroplast genome of Phalaenopsis aphrodite subsp.…”
Section: Sampling and Dna Extraction 152mentioning
confidence: 99%