Amplifikasi Gen COI dan 16s rRNA dari Invertebrata Laut Plakobranchus ocellatus
Abstract
ABSTRAK
Studi mengenai hubungan kekerabatan, evolusi dan identifikasi spesies secara molekuler tidak terlepas dari isolasi DNA dan amplifikasi gen COI dan 16s rRNA. Keberhasilan isolasi DNA dan amplifikasi gen COI dan 16S rRNA merupakan langkah awal yang penting untuk studi mengenai hubungan kekerabatan, evolusi dan identifikasi spesies. Penilitian ini bertujuan untuk mengamplifikasi gen COI dan 16S rRNA dari invertebrata laut yang diambil dari Raja Ampat, Plakobranchus ocellatus. Sepasang primer COI universal LCO-1490 dan HCO-2198 digunakan untuk mengamplifikasi fragmen gen COI dari P.ocellatus, sedangkan amplifikasi gen 16s rRNA menggunakan sepasang primer 16Sar-L dan 16Sbr-L. Fragmen gen COI dan 16S rRNA berhasil teramplifikasi dengan panjang amplikon masing-masing sekitar 680 bp dan 450 bp.
Â
Kata kunci: COI, gen, identifikasi, Plakobranchus ocellatus dan 16S rRNA
ABSTRACT
The first important step to study about organism relationship, evolution and species identification are DNA isolation and amplification of 16S rRNA and COI genes. This research aimed to amplify COI and 16S rRNA genes of marine invertebrates taken from Raja Ampat, Plakobranchus ocellatus. A pair of COI universal primer LCO-1490 and HCO-2198 was used for ampification a fragment COI gene P.ocellatus, while a fragment of the 16s rRNA gene was amplified using 16Sbr-L and 16Sar-L primer. Either  a fragment COI or 16S rRNA genes was successfully amplified, with a length amplicon about 680 bp and 450 bp, respectively.
Keywords : COI, gene, identification, Plakobrancus ocellatus, and 16S rRNA
References
Feng, Y., Q. Li, L. Kong, dan X. Zheng. 2011. DNA barcoding and phylogenetic analysis of Pectinidae (Mollusca: Bivalvia) based on mitochondrial COI and 16S rRNA genes. Mol Biol Rep (2011) 38:291–299.
Hebert, P.D.N., A. Cywinska, S. L. Ball, & J.R. deWaard. 2003. Biological identifications through DNA barcodes. Proc. R. Soc. Lond. B. Biol. Sci. 270, 313–321.
Hajibabaei, M., M.A. Smith, D.H. Janzen, J.J Rodriguez, J.B Whitfield, & P.D.N. Hebert. 2006. A minimalist barcode can identify a specimen whose DNA is degraded. Mol Ecol Notes 6:959–964.
Guo, X.D., D. Chen, T. J. Papenfuss, N.B. Ananjeva, D.A. Melnikov, Y. Wang. 2011 Phylogeny and divergence times of some racerunner lizards (Lacertidae: Eremias) inferred from mitochondrial 16S rRNA gene segments. Molecular Phylogenetics and Evolution 61 (2011) 400–412.
Vences, M., M. Thomas, A. van der Meijden, Y.Chiari dan D.R Vieites. 2005. Comparative performance of the 16S rRNA gene in DNA barcoding of amphibians. Frontiers in Zoology 2005, 2 :5.
Christa, G., L. Wescott, T.F. Schaberle, G.M. Konig, & H. Wagele. 2012. What remains after 2 months of starvation? Analysis of sequestered algae in a photosynthetic slug, Plakobranchus ocellatus (Sacoglossa, Opisthobranchia), by barcoding. Planta.
Ni, L., Q. Li, L. Kong, S. Huang, dan L. Li. 2012. DNA barcoding and phylogeny in the family Mactridae (Bivalvia:Heterodonta): Evidence for cryptic species. Biochemical Systematics and Ecology 44 (2012) 164-172
Takano, T., Y.M. Hirano, C.D. Trowbridge, Y.J. Hirano dan Yasuyuki Watano. 2013. Taxonomic clarification in the genus Elysia (Gastropoda: Sacoglossa): E. atroviridis and E. setoensis. Amer. Malac. Bull. 31(1): 25–37 (2013).
Folmer, O., M. Black, W. Hoeh, R. Lutz, & R. Vrijenhoek. 1994. DNA primers for amplification of mitochondrial Cytochrome c Oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology (1994) 3(5), 294-299..
Palumbi, S., A. Martin, S. Romano, W. O. McMillan, L. Stice, dan G. Grabowski. 1991. The Simple Fool’s Guide to PCR. Version 2.0. Department of Zoology and Kewalo Marine Laboratory, University of Hawaii, Honolulu, Hawaii.`
Downloads
Published
Issue
Section
License
Copyright and Attribution:
Articles in Biotropika: Journal of Tropical Biology are under Creative Commons Attribution (CC-BY-SA) copyright. The work has not been published before (except in the form of an abstract or part of a published lecture or thesis) and it is not under consideration for publication elsewhere. When the manuscript is accepted for publication in this journal, the authors agree to the automatic transfer of the copyright to the publisher.
Journal of Biotropika: Journal of Tropical Biology is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Permissions:
Authors wishing to include figures, tables, or text passages that have already been published elsewhere and by other authors are required to obtain permission from the copyright owner(s) for both the print and online format and to include evidence that such permission has been granted when submitting their papers. Any material received without such evidence will be assumed to originate from one of the authors.
Ethical matters:
Experiments with animals or involving human patients must have had prior approval from the appropriate ethics committee. A statement to this effect should be provided within the text at the appropriate place. Experiments involving plants or microorganisms taken from countries other than the author's own must have had the correct authorization for this exportation.