Optimization of the Promega PowerSeq™ Auto/Y system for efficient integration within a forensic DNA laboratory

Area of Science:

• Forensic Science
• Molecular Biology
• Genetics

Background:

• Massively parallel sequencing (MPS) is increasingly used in forensic DNA analysis for degraded or limited samples.
• Current MPS workflows require laborious library preparation, limiting practical application in casework.
• Forensic laboratories seek efficient methods to maximize information from challenging samples.

Purpose of the Study:

• To optimize the Promega PowerSeq™ Auto/Y System Prototype for forensic DNA casework.
• To introduce automation and reduce manual processing steps in MPS library preparation.
• To enhance laboratory efficiency and reliability for MPS implementation.

Main Methods:

• Automated bead-based purification replaced column-based PCR purification.
• A liquid handling robot platform was utilized for library preparation.
• Time-consuming quality checks were removed to streamline the workflow.

Main Results:

• The optimized workflow demonstrated concordance with capillary electrophoresis (CE) and standard MPS data.
• Key metrics like read abundance and allele balance remained consistent.
• Manual processing time was reduced by approximately 12 hours, increasing laboratory efficiency.

Conclusions:

• The optimized MPS workflow offers a more streamlined and efficient option for forensic DNA laboratories.
• Automation and reduced steps minimize contamination and human error risks.
• This improved MPS protocol facilitates practical implementation for forensic casework analysis.