PCR optimisation of newly designed complete mitochondrial DNA control region primers for species identification and genetic variants of the neglected gibbons of Sarawak

Renitha, Rajandran (2022) PCR optimisation of newly designed complete mitochondrial DNA control region primers for species identification and genetic variants of the neglected gibbons of Sarawak. [Final Year Project Report] (Unpublished)

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Abstract

Family Hylobatidae which includes gibbons and tiny apes, is the sister group to the Family Hominidae which includes great apes and humans. In Sarawak, two gibbon species exist; the Abbot’s Gray Gibbon (Hylobates abbotti) and Northern’ Gray Gibbon (Hylobates funereus), however, their evolutionary position is still unclear and there is a significant knowledge gap about these gibbon species particularly in terms of their phylogenetic relationship within the Family Hylobatidae. This study emphasizes on PCR optimisation of newly designed primers for species identification and phylogenetic analysis of the neglected gibbons of Sarawak. The blood samples of four captive individuals of H.abbotti and 1 individual of H.funereus were collected from Matang Wildlife Center, Kuching, Sarawak. Approximately 1092 base pairs of the control region segment of the mitochondrial DNA were analyzed using speciesspecific primers. The annealing temperature of the primers were optimized in the range between 45℃ to 60℃. The phylogenetic trees were constructed using MEGA 11 software for Neighbour-Joining (NJ), Maximum Likelihood (ML) and Maximum Parsimony (MP). This study results that the phylogenetic relationship of two individuals which were grouped together with H.abbotti sequences were solved whereas the phylogenetic relationship of other three individuals are still remain unresolved. This study also shows that although control region segment is phylogenetically informative, it is not able resolve the separation between H. abbotti and H.funereus species however, the segment is able to separate H.muelleri species from H.abbotti and H.funereus. In summary, the newly designed primers are suitable to detect the gibbon samples however the control region is not able to identify the positions between H.abbotti and H.funereus. Further studies are needed using a complete mitogenome in order to resolve the phylogentic positions of this gibbons’ species.

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