Simplified Direct PCR Method for Reference Buccal Samples Using a Non‐FTA Card by Omitting the Pretreatment Step. Issue 1 (21st August 2019) - Jung, Ju Yeon
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.