Genetic identification of stone crab (Myomenippe spp.) with DNA barcoding approach caught in Tarakan city sea waters

Stone Crab is a fishery potential obtained from the sea waters of Tarakan City that have not been utilized. These crabs have various morphological variations, so it is necessary to determine the molecular status of the species. Molecular identification is usually made using DNA barcoding techniques. DNA barcoding is a method to determine the taxonomy of organisms molecularly by using DNA chains to identify an organism quickly and accurately. This study aims to genetically identify stone crab samples obtained from the sea waters of Tarakan City and compare their sequences with the GenBank database. DNA extraction was carried out following the protocol of the DNA extraction kit (genomic DNA Purification wizard, Promega). COI gene fragments were amplified by PCR method, using COI Universal Primer. Sequencing was carried out in two directions, forward and reverse with the Sanger termination dideoxy method. The DNA sequencing process is carried out by first Base company in Singapore. The results of the DNA sequence obtained have a length of 599-832 base pairs. The results of the comparison of homology with the NCBI database and the BOLD system show that this sample has similarities with the COI sequences of Myomenippe fornasinii and Myomenippe hardwickii with sim,ilarities reaching 91.56% to 97.22%. The genetic distance obtained from the genetic distance analysis shows that samples C2 and C1 have the closest genetic distance value with a value of 0.019, and the farthest value obtained between samples is C1 and B2, which is 0.041. The results of the phylogenetic analysis showed the formation of 2 clusters. In that cluster, A was filled with samples A1, A2, B2, C1, and C2 and for cluster B was filled with additional data obtained from the NCBI GenBank.