With MPS, sequences can be analyzed more in depth to determine whether they are genuinely from one of the original contributors of a sample, or instead more likely to be the product of a PCR or sequencing error. Additionally, due to the ability to multiplex more loci than CE affords, broader genetic interrogation can be achieved in a single reaction, thus
conserving precious samples. The reported results comprise only 16 loci, but MyFLq can run with any number of loci. When running MyFLq with a custom loci set, the primers of these loci can be imported. The allele database is not strictly necessary to run the program. In exploratory studies, for example if building a database of known alleles, MyFLq can be run with an empty allele database. The GitHub repository contains example files for users that need either a custom RGFP966 solubility dmso locus set or custom allele database. The used allele database was very small as it only compromised the alleles of the five contributors. Sequences AZD2281 price that are currently not in the database are marked as red bars. These bars are very useful to visually monitor the noise level. In the future, with a larger database, it could be that erroneous sequences are nonetheless present in the database, as they could be true alleles for individuals that are not present in the sample. The solution to that problem could be to mark rare alleles (e.g. alleles with a population prevalence
<1%) with a different color. The combination
of unknown alleles and rare alleles would then indicate the level of noise. A further limitation of the current database is its nomenclature. Currently same-sized alleles get an arbitrary name within the mafosfamide database, which would make it difficult to perform searches in other databases without the original sequence. When an international nomenclature for MPS STR alleles has been established, it will be incorporated in MyFLq. When all allele candidates have been reviewed, the “Make profile” action generates a report with only the selected alleles. This is the profile that a forensic analyst can use to either store in a database, to query against a database, or for direct comparison to a known sample of interest. Future versions of the software will include possibilities to interact directly with sample databases. New feature requests can be made through the GitHub website. MyFLq is the first open-source, web-based forensic MPS DNA analysis software with an easy-to-use graphical user interface. It can run natively on Illumina BaseSpace, or independently on a forensic laboratory’s server. The possibility to run the program directly from the Illumina BaseSpace environment means no extensive bioinformatics skills are required. C.V.N. participated in an internship program at Illumina, Inc. to provide feedback on building a native BaseSpace application to the Illumina developers.