VARIATION OF MITOCHONDRIAL DNA D-LOOP REGION MUTATION ON HUMAN TISSUES FROM EMBRYO ECTODERM LAYER

• Main Author: MASYKURA (NIM 10505081), NAJMIATUL
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/12452
• Institution: Institut Teknologi Bandung

Fertilization process yield single cell with numerous copies of mitochondrial DNA (mtDNA). There is no information weather mutation happen in embryogenesis process until formation of three embryo layers; ectoderm, mesoderm, and endoderm layer. The aim of this research is identifying mutations in mtDNA D-loop region in tissues from ectoderm layer. Skin and oral mucosa tissue samples as the object of this research autopsied from one person. Each sample was lysised to obtain the DNA template, and amplified with Polymerase Chain Reaction (PCR) method. To obtain nucleotide sequences, DNA fragment from amplification process are sequenced with Sanger dideoxy method. Mutation in these samples identified by in silico analysis. After agarose gel electrophoresis, 0.9 kb mtDNA D-loop fragmen band was observed. We obtain the sequence of 932 nucleotide base from electroforegram. In silico analysis show some nucleotide differences between samples and revised Cambridge Reference Sequence (rCRS), in position and type. There are some mutation differences between both samples. Based on this result, we know that the D-loop mtDNA nucleotide sequences in both sample are not identical. We recommend further research about mtDNA nucleotide sequences homology of several tissues from one person. Mitochondrial DNA nucleotide sequence differences among tissues in one person suggest the using of the same tissue type as mtDNA template in forensic analysis.