Simplified Direct PCR Method for Reference Buccal Samples Using a Non-FTA Card by Omitting the Pretreatment Step

Area of Science:

• Forensic Science
• Molecular Biology
• Genetics

Background:

• Non-FTA cards are commonly used for biological sample collection.
• Direct PCR amplification from non-FTA cards often requires pretreatment steps with specific buffers.
• Optimizing direct PCR protocols is crucial for efficient forensic DNA analysis.

Purpose of the Study:

• To develop and validate a rapid direct PCR method using a non-FTA card (Oral Cell Sampling Kit).
• To assess the compatibility of this method with commercial multiplex STR kits.
• To optimize PCR conditions for high-throughput analysis of Korean buccal samples.

Main Methods:

• A direct PCR method was designed, omitting the Prep-n-Go™ Buffer pretreatment step for non-FTA cards.
• The method's compatibility was tested with GlobalFiler™ Express PCR Amplification Kit, GlobalFiler™ PCR Amplification Kit, and PowerPlex® Fusion system.
• PCR conditions, including final volume and cycles, were optimized.
• Performance was evaluated using 50 Korean buccal samples.

Main Results:

• The developed direct PCR method demonstrated compatibility with GlobalFiler™ Express, GlobalFiler™, and PowerPlex® Fusion systems.
• Optimized conditions involved a 10 µL final PCR volume and 29 cycles.
• Over 90% of samples yielded full profiles when using GlobalFiler™ PCR Amplification Kit and PowerPlex® Fusion system.
• The method proved effective for analyzing Korean buccal samples.

Conclusions:

• A simplified, rapid direct PCR protocol for non-FTA cards was successfully established.
• This method eliminates the need for pretreatment buffers, streamlining sample analysis.
• The protocol is suitable for forensic applications, particularly for analyzing Korean reference samples collected on non-FTA cards.