NUCLEOTIDE SEQUENCE OF MITOCHONDRIAL DNA D-LOOP FRAGMENTS IN TISSUES FROM MESODERM LAYER

• Main Author: NaulinA Simarmata, Eva
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/35120
• Institution: Institut Teknologi Bandung

Embryogenesis leads formation of three embryonic layers. Those are endoderm, mesoderm, and ectoderm. All layers will be differentiated into specific organs or tissues. During that process, information about nucleotide changing in mitochondrial DNA (mtDNA) of the D-loop fragment has not been investigated. The aim of this study is to know whether there is any nucleotide changing between psoas and diaphragm muscle that come from the mesoderm layer. To achieve the research objective, cell lysis, DNA amplification using Polymerase Chain Reaction (PCR), agarose gel electrophoresis, sequencing, and in silico data analysis were used. PCR result showed a 0.9 kb ribbon in agarose gel electrophoresis. Sequencing result, called electrophoregram, showed that psoas has 918 bp and diaphragm has 892 bp of nucleotides. In silico results showed both of psoas and diaphragm share identical nucleotide sequences. Furthermore, nucleotide sequence comparisons between two samples and revised Cambridge Reference Sequence (rCRS) showed ten mutations. One of the ten mutations has not been published yet, that is C(253)T. These results should be explored further for forensic purposes. The forensic analysis will take advantages of the same nucleotides sequences of samples that come from one embryonic layer. Tissues from one embryonic layer can be used as a source of DNA template.