SANGER SEQUENCING FOR MOLECULAR KINSHIP DETERMINATION IN IRAQI FAMILIES WITH MISSING PATERNAL MEMBERS

Abstract

Background: Kinship testing is crucial during forensic investigations into missing persons, and Sanger sequencing has proven to be an effective method for establishing biological connections. The present work aimed to establish the worth of Sanger sequencing to resolve paternity claims among Iraqi families whose fathers have gone missing.

Materials and Methods:  Genomic DNA was extracted from 300µl of whole blood, collected from six cases (n=120 samples) at the Medical Legal Directorate in Baghdad, Iraq. Each case comprised three relatives of the missing father, the mother, and the putative child. Polymerase Chain Reaction (PCR) procedures were performed using specific primers for the D18S, TPOX, CSF, and FGA genes. Sanger sequencing was utilised to compare the sequences from the relatives with those of the child.

Results: The findings of the present study revealed the sequence of outcomes for six Iraqi cases of missing fathers. The range of DNA concentration was (75 to 110 ng/µL). The findings revealed a high degree of sequence similarity between the grandson and his grandparents, as well as among the siblings, aligning with the expected patterns of inheritance. The sequencing data for six families revealed the matching alleles used in the study for each family member, which supported the pedigree hypothesis and confirmed the biological connections between the grandparents, siblings, and the grandson.

Conclusion: In conclusion, Sanger sequencing is widely utilised in kinship analysis due to its accuracy, reliability, and specific application scope. In certain situations, such as cases involving a missing father where traditional methods fall short, it is considered a crucial method for confirming paternity alongside other techniques like Short Tandem Repeats.