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Simplified Direct PCR Method for Reference Buccal Samples Using a Non-FTA Card by Omitting the Pretreatment Step
Authors:Ju Yeon Jung M.S.  Ga Hee Ryu B.S.  Da-Hye Kim B.S.  Joo-Young Kim Ph.D.  In Kwan Hwang Ph.D.  Pil-Won Kang Ph.D.  Byung Won Chun Ph.D.  Seong Yeon Yoo Ph.D.
Affiliation:Forensic DNA Division, National Forensic Service, Wonju, 26460 South Korea
Abstract:When using non-FTA cards in commercial multiplex STR kits for direct PCR, pretreatment steps with specific buffers are recommended. Here, we designed a rapid direct PCR method utilizing a non-FTA card, Oral Cell Sampling Kit, by omitting the pretreatment step involving Prep-n-Go™ Buffer, and it showed compatibility with the GlobalFiler™ Express PCR Amplification Kit, GlobalFiler™ PCR Amplification Kit, and PowerPlex® Fusion system. To optimize the PCR conditions, we tested the method with different final PCR volumes and cycles. Finally, we conducted a performance test using 50 Korean buccal samples and confirmed the high performance of the method, detecting more than 90% of the samples with full profiles when using GlobalFiler™ PCR Amplification Kit and PowerPlex® Fusion system at 29 cycles in a 10 μL final PCR volume. Thus, we report a simple direct PCR set-up to analyze reference samples collected using a non-FTA card manufactured in Korea.
Keywords:forensic science  DNA typing  direct polymerase chain reaction  Oral Cell Sampling Kit  Prep-n-Go™ Buffer  reference samples
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