Development of forensic genomics research toolkits by the use of Massively Parallel Sequencing

Abstract

Currently, Forensic DNA research is conducted almost exclusively using capillary electrophoresis to determine the length of fragments containing Short Tandem Repeats (STRs). Over the past decade, developments in MPS (Massively Parallel Sequencing techniques, also known as Next Generation Sequencing) offered new possibilities for forensic DNA research. This thesis focusses on the preparation, validation and implementation of MPS and the accompanying data analysis. By MPS, the exact DNA sequence of STRs is determined often revealing additional variation on top of the fragment length resulting in an even more unique DNA profile. Sequence information provides more insight on the DNA molecules comprising a conventional DNA profile. With this information, a software could be developed to make a better distinction between genuine alleles and noise offering possibilities for analysis of unbalanced DNA mixtures which are often encountered in forensic casework.As alternative DNA marker to STRs, research was conducted to select microhaplotypes (multiple variable positions in a small fragment). The statistical power of a profile generated from these microhaplotypes turned out to be almost as strong as that of STRs without suffering from the known STR-artefacts thereby offering possibilities for interpretation of DNA mixtures. </p