Assay Development and Validation of an 8-SNP Multiplex Test to Predict Eye and Skin Coloration

Area of Science:

• Forensic genetics
• Human identification
• Phenotypic prediction

Background:

• Accurate identification of human remains is crucial in forensic science.
• Predicting physical traits like eye and skin color from genetic data aids identification.
• Existing methods may not be robust for challenging forensic samples.

Purpose of the Study:

• To develop and validate a multiplex assay for predicting eye and skin color using single nucleotide polymorphisms (SNPs).
• To assess the assay's performance on various forensic samples, including degraded DNA.

Main Methods:

• Development of an 8-SNP multiplex assay utilizing multiplex PCR and single-base primer extension.
• Detection of fluorescently labeled oligonucleotides via multicolor capillary electrophoresis.
• Validation included tests for reproducibility, reliability, sensitivity, species specificity, and performance on degraded and forensic DNA samples.

Main Results:

• The 8-SNP multiplex assay demonstrated robustness and reliability across validation tests.
• The assay successfully performed on challenging forensic samples, including bone.
• Genotypes were reliably determined for predicting eye and skin color.

Conclusions:

• The developed 8-SNP multiplex assay is a valuable tool for forensic geneticists.
• This assay can reliably predict eye and skin color from challenging DNA samples.
• The assay significantly aids in the identification of human remains and missing persons.