Phylogeny reconstruction of Gerridae from River Ratatotok and River Talawaan North Sulawesi Based on Mitochondrial DNA 16 S RNA Gene

Abstract

Has conducted research that aims to get the position species based Gen 16 S RNA Mitochondrial DNA of Water Striders in Talawaan River and Ratatotok River, North Sulawesi. Extraction of total DNA using DNA Innuprep Micro KIT (Analytic Jena, Germany). 16 S RNA gene [16sA (CTG 5'CGC AAA TTT AAC AAC AT 3'); 16sB2 (5'TTT AAT ACA TCG AGG CCA 3')]. 16 S RNA gene amplification using Master Cyler with Rotor Gene Q Series Software 2.0.3. PCR conditions that denaturation initiation stage 94 0C for 3 minutes, denaturation stage 94 0C for 30 seconds, annealing step 52 0C for 50 seconds, extension stage 72 0C for 1 min and final extension 72 0C for 5 minutes. Visualization amplicon using Automatic Electrophoresis Qiaxel. Sequencing was performed using the ABI PRISM 3730xl Genetic Analyzer Develop by Applied Biosystems, USA. Construction phylogeny using MEGA 6.0 program. Based on the phylogeny tree formed from Talawaan Water striders have closest kinship with Limnoporus esakii while Water striders of Ratatotok has the closest kinship with Cylindrostethus costalis.