Development of human identification kit using nanopore sequencing technology

Objective: Human DNA is one of the most stable biological materials known and contains a number of individual identification sites. In recent years, genotyping of certain SNP on DNA has become increasingly common, especially in identification studies, and offers high discrimination power. Although the genotyping process has become increasingly practical due to the rapid advances in next generation sequencing technology, many of these devices have long-lasting results and require large data analyzes, especially in cases where rapid identification is required. However, the Minion device, which is defined as the 4th generation sequence device, has a small size and enables rapid analysis, thus eliminating these disadvantages. In this thesis; it is aimed to develop a technological and practical SNP based human identification prototype that will adapt to the Oxford Nanopore sequencing platform. Materials and methods: Using commercial DNA, 52 SNP regions frequently used for identification in the literature were amplified by multiplex PCR method. These amplicons were sequenced with the Oxford Nanopore Minion device. A unique bioinformatics workflow is designed based on sequence data. Results: Nanopore data were found to be completely accurate in 52 SNP regions in three commercial DNA samples. Total time was completed under 1 hour. Conclusion: As a result of this thesis, 52 SNP sites were sequenced successfully with Oxford Nanopore Minion device. At the same time, the fastest time has been achieved in the world for SNP based human identification.