ANALYSIS METHODE OF 917 BASE PAIR IN D-LOOP HUMAN MTDNA

• Main Author: RACHMADI, BAMBANG
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/5830
• Institution: Institut Teknologi Bandung

Genetic mitochondrial DNA ( mtDNA ) has a high mutation rate. This mutation pattern can be<br /><br /> used for forensic, evolution and genealogy study, particularly the D-loop region, consist of<br /><br /> hypervariable segment 1 (HVS1) and hypervariable segment 2 (HVS2). Research the D-loop<br /><br /> region begin with amplification and sequencing until now is done separately, requiring a longer<br /><br /> time to analysis. Based on that reason, development on the method to analyze this region is<br /><br /> important. In this research, 0.9 kb DNA fragment in Human D-loop mtDNA was amplified with<br /><br /> Polymerase Chain Reaction (PCR). This research begin with lysis to obtain DNA template, then<br /><br /> it was amplified using Polymerase Chain Reaction (PCR) method, followed by sequencing of<br /><br /> the amplified mtDNA using Dideoxy Sanger method to confirm, along with nucleotides analysis<br /><br /> using SeqmanTM version 4.0.0. The result of this research shows 0.9 kb single band on the<br /><br /> agarose gel electrophoresis. Electrophoregraph data shows the 0,9 kb nucleotides sequent is<br /><br /> inside the D-loop region and demonstrated 8 mutations after nucleotide sequence compare with<br /><br /> Cambridge Reference Sequence (CRS). The result of this method, gives amplification on the<br /><br /> region of HVI-HVII could be done in one step reaction, so analysis for D-loop region is more<br /><br /> effective and efficient.