Isolation and Characterization of a Partial Fragment of Mitochondrial Genome (tRNA-Gln' -tRNA-Lys') of Rasbora sarawakensis

Lim, Woan Ting (2017) Isolation and Characterization of a Partial Fragment of Mitochondrial Genome (tRNA-Gln' -tRNA-Lys') of Rasbora sarawakensis. [Final Year Project Report] (Unpublished)

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Abstract

Rauhora sarax akensis is well known as Sarawak Rasbora that belongs to family Cvprinidae. This fish is significant for ornamental purposes and as molecular tools to study the phylogenetic relationship among the other Rasbora spp. In order to determine the phylogenetic relationship, mitochondrial genome is being targeted for analysis. In this study, our purpose is to isolate and characterize a partial fragment of mitochondrial genome (tRNA-GIn'-tRNA- /. vs') of R. saraxnkensis. The collection of R. sarawakensis was conducted at Matang Wildlife Park and acclimatized at 26 °C, 12 hours dark and light photoperiod. DNA extraction was conducted using CTAB method and the fragment of mitochondrial genome was amplified through PCR. Four pairs of gene specific primer were designed for PCR amplification of this fragment based on the conserved sequences from multiple alignment of three selected freshwater fishes. PCR amplification had generated four fragments of PCR amplicons in which their length are close to the expected size during primer design. The PCR products were purified using QlAquick PCR Purification Kit (Qiagen, Germany) and was analyzed through AGE. Subsequently, the sample was sent to the First BASE Company for sequencing. The E-value and blast identity obtained for all of the four fragments are 0.0 and more than 80% respectively. Sequence that is obtained was aligned to other previously isolated fragment to generate complete mitochondrial fragment for R. sarawakensis. Phylogenetic analysis revealed high bootstrap support between R. sarmrcrkensis and other closely related Rasbora spp. This study serves as a platform for further understanding in the population genetic of Rauhora spp. that might be useful in future conservation effort.

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