Report of Systematic Zoology Lab Practicum, Volume 4: e17; August, 2013


An attempt to extract DNA from the spiny sand seastar Luidia quinaria (Echinodermata: Asteroidea) collected in Oshoro Bay, Hokkaido, northern Japan


Yasushi Isozaki and Naho Yagi

Division of Biology, Department of Biological Sciences, School of Science, Hokkaido University, Sapporo 060-0810, Japan



Material and Methods
This star fish was obtained at Oshoro Bay, Hokkaido, Japan, about 43°12′N, 140°51′E, on 12 June 2013 by Yasushi Isozaki and Naho Yagi, photographed and identified by Hiroshi Kajihara as Luidia quinaria by a reference to Oguro (1992), and fixed in 99% EtOH. We tried to extract DNA from a tube foot using the silica method (Boom et al. 1990) with some modifications. Remaining morphological voucher specimen has been deposited at the Hokkaido University Museum under the catalogue number ICHU2112063 (contact: Dr. Hiroshi Kajihara, kazi@mail.sci.hokudai.ac.jp).
      None of the attempts were sucessful to PCR amplify the mitochondrial cytochrome c oxidase subunit I gene (COI) using LCO1490 (5′-GGTCAACAAATCATAAAGATATTGG-3′) and HCO2198 (5′-TAAACTTCAGGGTGACCAAAAAATCA-3′) (Folmer et al. 1994) and the 28S rRNA gene using LSU5 (5′-ACCCGCTGAAYTTAAGCA-3′) and LSU3 (5′-TCCTGAGGGAAACTTCGG-3′) (Littlewood et al. 1994).

Taxonomy
Phylum Echinodermata
Subphylum Asterozoa
Class Asteroidea
Superorder Valvatacea
Order Paxillosida
Family Luidiidae
Genus Luidia
Luidia quinaria Von Martens, 1865
[Japanese name: suna-hitode]
(Figs 1, 2)



Fig. 1. Luidia quinaria (ICHU2112063), aboral view.


Fig. 2. Luidia quinaria (ICHU2112063), oral view.



References

Boom, R., Sol, C. J. A., Salimans, M. M. M., Jansen, C. L., Wertheim-van Dillean, P. M. E., and van der Noordaa, Jand Dixon, J. 1990. Rapid and simple method for purification of nucleic acids. Journal of Clinical Microbiology 28: 495–503.

Former, O., Black, M.Hoeh, W., Lutz, R. 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3: 294: 294-299.

Littlewood, D. T. 1994. Molecular phylogenetics ofcupped oysters based on partial 28S rRNA gene sequences. Molecular Phylogenetics and Evolution 3: 221–229.

Oguro, C. 1995. Asteroidea. Pp. 513–529. In: Nishimura, S. (Ed.) Guide to Seashore Animals of Japan with Color Pictures and Keys, Vol. II. Hoikusha, Osaka.