Low Volume STR Amplification Options: Coupling with Standard or Fast PCR, Traditional or Normalized DNA Extraction,...

Area of Science:

• Forensic Science
• Molecular Biology
• Genetics

Background:

• Short Tandem Repeat (STR) amplification is crucial for forensic DNA profiling and human identification.
• Current amplification methods can be costly, prompting the need for cost-saving strategies.
• Reduced reaction volumes offer a viable cost-saving measure, particularly for high-quality DNA samples.

Purpose of the Study:

• To present optimized protocols for reduced-volume STR amplification (3µL and 6µL) for forensic applications.
• To evaluate the use of commercial amplification kits with buccal samples.
• To detail methods for DNA extraction and capillary electrophoresis detection compatible with reduced reaction volumes.

Main Methods:

• Utilized 3µL and 6µL reaction volumes with commercial amplification kits (KAPA2G™ Multiplex Mix) for buccal samples.
• Employed standard and fast polymerase chain reaction (PCR) protocols.
• Integrated normalized DNA extraction using the ChargeSwitch® Forensic DNA Purification Kit.
• Applied capillary electrophoresis (CE) detection using POP-4™ polymer with a 36cm array and POP-6™ polymer with a 22cm array on 3130 Genetic Analyzer instruments.

Main Results:

• Demonstrated successful STR profile development using reduced-volume reactions for buccal samples.
• Showcased compatibility with both traditional and normalized DNA extracts.
• Validated alternative capillary electrophoresis detection methods for enhanced throughput or specific applications.

Conclusions:

• Reduced-volume STR amplification protocols are effective for forensic human identification using buccal samples.
• Optimized extraction and detection methods support cost-effective DNA profiling.
• These findings offer practical solutions for forensic laboratories seeking to improve efficiency and reduce costs.