Bode Technology™ Buccal DNA Collector™ designed to be easy to use and to store the sample. The main objective of this evaluation is to optimize a reduced volume PCR amplification for STR analysis of reference buccal samples collected on Bode Technology™ Buccal DNA Collector™. A total number of 21 buccal samples collected on Bode Technology™ Buccal DNA Collector™ were used for this evaluation. A 1.2 mm disc was punched from the samples into a 96-well plate using BSD600-Duet Punching System. The samples were amplified using 14.5 μl, 10 μl and 8 μl volume of GlobalfilerTM Express amplification mix at 25 cycles with a GeneAmp PCR System 9700. The electrphoresis of PCR products was carried out on ABI 3500xL Genetic Analyzer. An electrophoresis master mix was prepared for each sample using 9.5 μl of Hi-Di™ formamide and 0.5 μlGeneScan™ 600 LIZ® size standard. After that, 1 μl of PCR product was used from each sample. Data analysis using GeneMapperID-X Software v1.4. Correct profiles were observed for all samples. One sample produced no profile when using 8 μL of amplification master mix. No partial profiles were observed when amplified with 8 μL, 10 μL and 14.5 μL of the amplification mix. All amplification mix volumes showed off-scale results for several samples. Reducing the amplification mix volume to 8 μl for PCR amplification using Globalfiler™ Express assay, resulted in a first-pass success rate of 95.2%. This was slightly lower than 10 μl and the full volume amplification mix. First-pass success rate with 10 μl of amplification mix for direct amplification achieved 100%. The volume using 10 μl amplification mix was found to be reliable to generate full profile. This approach can reduce the cost of DNA profiling of buccal samples on Buccal DNA Collector™ using GlobalfilerTM Express assay.
Direct PCR amplification,buccal samples, Buccal DNA Collector.