From Janosik et al (2013):
Specimen collection
Specimens were obtained from the Division of Echinoderms, Smithsonian Institution National Museum of Natural History (USNM) in Washington, DC, the Department of Invertebrate Zoology, California Academy of Sciences (CASIZ), San Francisco, California, and the National Institute of Water and Atmospheric Research (NIWA), New Zealand (Table 1). Most specimens were dried. Antarctic species were collected during two five-week research cruises aboard the R/V Laurence M. Gould in November/December of 2004 and May/June of 2006. Images of D. clarki were provided by NIWA.
Molecular data
Molecular methods follow Janosik et al. (2011). DNA extraction of specimens was performed by using DNeasy Tissue Kit (Qiagen). Two mitochondrial DNA markers (16S and COI) were utilized to estimate the evolutionary history of Odontasteridae. Specifically, a 508-bp region of the mitochondrial 16S gene was amplified using the 16SarL and 16SbrH primers and protocols of Palumbi et al. (1991). For the same individuals, a 627-bp region of the COI gene was amplified using primers designed to work with Odontaster COI-Ast 22F (5'-TTYTCNACNAAACA YAAGGA-3') and COI-Ast722R (5'-GGRTGNCCRAAR AAYCARAA-3') (Janosik et al. 2011). Amplified products were purified with either a Qiagen QIAquick Gel Extraction Kit (Qiagen Inc.) or a Montage PCR Filter Units (Millipore) according to the manufacturer’s directions. Purified products were then sequenced bidirectionally on a Beckman CEQ 8000 Genetic Analysis System (Beckman Coulter). Sequences were edited and aligned using Sequencher 4.6 (Gene CodesCorporation) andBioedit v.7.0.8 (Hall 1999). COI sequences were translated according to the echinoderm mitochondrial DNA code to aid in proofreading. Genbank accession number is listed in Table 2.
Based on current understandings of sea star relationships (Blake 1987; Mah and Foltz 2011), Bathybiaster loripes, Chaetaster moorei, Crossaster papposus, Luidia foliolata, Mediaster aequalis, and Solaster stimpsoni were selected for the outgroup, and these sequences were downloaded from GenBank (www.ncbi.nlm.nih.gov/ genbank/) (Table 2).