VARIAN NORMAL FRAGMEN 0,4 KB DAERAH D-LOOP DNA MITOKONDRIA MANUSIA INDONESIA

• Main Author: Gaffar, Shabarni
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/4729
• Institution: Institut Teknologi Bandung

<b>Abstract:</b><p align=\"justify\"> <br /> A good standard reference for the highly polymorphic human mtDNA sequence is essential for studies of normal variant and desease-related nucleotides variant in the mitochondrial genome. However, the published reference data base was build from mtDNA sequence of Caucasian origins. As already observed that sequence polymorphism of Indonesian mtDNA exist, this major research aims to build normal variant data base of the Indonesian human mtDNA and translational products, the specific aim is to determine normal variant of 0.4 kb D-loop Indonesian mtDNA. To obtain those goals, ten samples from Batak, Palembang, Badui Sundanese, Javanese, Balinese, Bima, Banjarmasin, Gorontalo and Temate were used as sources of mtDNA. Template DNAs which were lysed from ephitelial cells were amplified by PCR method employing Ml dan M2 primers. PCR product then was ligated to T-pMOSBlue vector and cloned into E, coil MOSBIue. Sequencing of 0.4 kb D-loop mtDNA were carried out by dideoxy Sanger Dye terminator method. The result of PCR process showed one band of 0.4 kb DNA fragment through agarose gel electrophoresis. This research also yielded 49 white colonies which contained insert DNAs. The result of sequencing of recombinant plasmid revealed that the size of the insert DNAs were either 443 or 445 bp. The total number of nucleotides sequenced was 4436 bp. Sequencing analysis subsequently showed that there were polymorphisms in 0.4 kb D-loop mtDNA. Nucleotide comparison analysis to the Anderson sequence (Cambridge sequence) showed that there were 24 nucleotide variants (morphs) in 0,4 kb D-loop mtDNA. Twenty three of 24 morphs have been already published. One of 24 morphs which was from Badui samples has never been published. Those morph presumably was a specific one for Badui sample, which identified at 16.109, A in Anderson sequence and G in Badui.<p align=\"justify\">