Performance comparison of a previously validated microhaplotype panel and a forensic STR panel for DNA mixture analysis.

Short Tandem Repeats (STRs) are the most widespread markers in forensic genetics. However, STR stutter peaks can mask alleles from a minor contributor when analysing mixtures, hindering the interpretation of complex profiles. In this study we compared the performance of a previously described panel of microhaplotypes (MHs), an alternative type of forensic marker, against a standard STR kit.

Exploring legal age estimation using DNA methylation.

Minors (subjects under the legal age, established at this study at 18 years) benefit from a series of legal rights created to protect them and guarantee their welfare. However, throughout the world there are many minors who have no way to prove they are underaged, leading to a great interest in predicting legal age with the highest possible accuracy. Current methods, mainly involving X-ray analysis, are highly invasive, so new methods to predict legal age are being studied, such as DNA methylation.

Adapting an established Ampliseq microhaplotype panel to nanopore sequencing through direct PCR.

We have adapted an established Ampliseq microhaplotype panel for nanopore sequencing with the Oxford Nanopore Technologies (ONT) system, as a cost-effective and highly scalable solution for forensic genetics applications. For this purpose, we designed a protocol combining direct PCR amplification from unextracted DNA with ONT library construction and sequencing using the MinION device and workflow. The analysis of reference samples at input amounts of 5-10 ng of DNA demonstrates stable coverage patterns, allele balance, and strand bias, reaching profile completeness and concordance rates of ∼95%.