Perpustakaan Digital ITB

Embryogenesis is initiated when oocyte fused with spermatozoon, each of them is from the reproductive cells. The zygote which formed will be splitted up to three embryonic layers, i.e. ectoderm, mesoderm, and endoderm. Until this day, there has not been reported whether differences in nucleotide sequence in the embryonic layer that can contributes in the field of forensics. The purpose of this research is to analyze the homology of the nucleotide sequence of mtDNA in various tissues and endoderm layers. The approach taken is using the Polymerase Chain Reaction (PCR) to amplify the mtDNA D-Loop. Nucleotide sequence of PCR results were determined by Sanger dideoxy method mtDNA. The origin of sample is from the endoderm layer, comes from the small intestine and pancreas are derived from a single individual. The results showed single band amplification on each sample with a size of approximately 0.9 kb on agarose gel electrophoresis. Electrophoregram data were obtained along the length of 906 bases for the small intestine and 825 base pairs for pancreas. Results of analysis in silico showed the same nucleotide sequence from both samples. The comparison of samples with the revised Cambridge Reference Sequence (rCRS) shows there are 13 differences in nucleotide sequence with two nucleotide sequence differences, that have not been reported, they are C132T and C253T. Based on our last research groups, there are no differences in nucleotide sequence between samples in the mesoderm layer of the same individual but not for three embryonic layers from different individuals. This indicates there are no mutation during embryonic process on a some individual. So the samples from the difference tissue can be used for forensic purposes.