DIFFERENCE OF HUMAN MTDNA SEQUENCE AT D-LOOP REGION ON TISSUES FORMED FROM MESODERM AND ENDODERM LAYER

• Main Author: YUNIARTI (NIM 10505017), NIA
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/12463
• Institution: Institut Teknologi Bandung

Fertilization leads to the formation of zygote which has mitochondrial DNA (mtDNA) with specific sequence. Following fertilization, the zygote undergoes several steps to become an embryo and this process includes the formation of three germ layers, ectoderm, mesoderm and endoderm. It has not been known already, if there is any difference of mtDNA sequence on each germ layers. Aim of this research is to see difference of nucleotide sequence in D-Loop region of mtDNA from tissues that formed from mesoderm and endoderm layer, compare with mtDNA sequence of Cambridge Reference Sequence (CRS). Sample that analyzed was heart and liver tissue from one individu. Lysis cell was done to get the mtDNA and it continued with amplification process of mtDNA using Polymerase Chain Reaction (PCR) method. Confirmation about PCR results used agarose gel electrophoresis method. The nucleotide sequence of mtDNA was known through DNA sequencing process by dideoxy Sanger's method. Experimental result showed the single band with 0.9 kb in size and electrophoregram result of DNA sequencing contained with 918 base pairs. In silico analysis showed differences sequence of mtDNA in D-Loop region between tissues from mesoderm and endoderm layer compared with Cambridge Reference Sequence (CRS). This result is new invention and is not reported yet. It opens wide range to other researchers to do further research about this case, and in maternal inheritance analysis using mtDNA, it is suggested to use cell from the same tissue. <br />