Abstract
Optimization and validation of direct amplification protocols for both AmpF lSTR ® Identifiler ® Direct (IDD) and PowerPlex ® 16 HS (PP16HS) on the high-throughput 3730 DNA Analyzer (Applied Biosystems) were carried out in preparation for potential changes in Canadian legislation. Amplification conditions were optimized to perform direct amplification of single source blood, buccal and extracted hair roots collected from convicted offenders (CO) on FTA ® paper without the need for sample purification prior to amplification. Validation was performed using IDD and PP16HS on a wide collection of blood and buccal samples from volunteer donors. In addition, 111 CO blood samples were selected to assess the robustness of the new analytical process (i.e. 0.53 mm FTA disks amplified in 10 μL PCR volume) and verify the stability of DNA stored on FTA paper at room temperature for up to 10 years. Studies such as reproducibility, robustness and concordance were carried out. The quality of the STR profiles generated using the new analytical process was assessed by examining profile intensity, heterozygote peak height ratios and interlocus balance. Reliable and high quality profiles from all CO samples and volunteer samples were produced under the new process giving support to its implementation.
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