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Abstrak Phylogenetic Analysis of Frogs in Kumbe River Based on COI Gene of Mitochondrial DNA

Chapter 2017 with 263 download times

Authors

,Arif Wibowo,Yoga Candra Ditya,

The level of genetic variation and population originallity is important for directing conservation and managemen purposes. Thus we are conducting the research to determine and discuss phylogenetic relationship of common frog in Kumbe River, Papua and also to pointed the possible acts of conservation of such aquatic resources in a riverine system. The specimens in the form of larval for the study were obtained from Kumbe River System, Papua on the year of 2014.  Total  genomic  DNA  was  extracted  from  whole larvae using  a  DNA  extraction  kit procedure and     a partial fragment of the mitochondrial COI was amplified using universal primers  Fish-COI-F  and  COI-Fish-R.  The  PCR  products  were  visualised  and  sequencing reaction was performed by the EZ-Seq service (Macrogen) using the reverse primer (COI-Fish- R). Chromatograms were checked manually and multisequence alignments were done using MUSCLE. A neighbour-joining tree was constructed with the Kimura 2-parameter (K2P) model using MEGA version 5.0 software to infer the phylogeny of this species. The results reveal that PCR products of the Cytochrome Oxidase Subunit I (COI) mtDNA regions for the three common frog ponds samples produced fragments of 570 bp. DNA barcoding unable the species identification of larval morph types and the minimum genetic distance (0.171) or 82.9% similarities to the closes species. However the sequence of mitochondrial DNA gen COI has been successfully used in phylogeny analysis. The samples collected were identified only on genus level Rana sp, which has unique characteristic of nucleotide composition.

Key words : Kumbe, phylogenetic,pond, frog