Jung Ju Yeon, Ryu Ga Hee, Kim Da-Hye, Kim Joo-Young, Hwang In Kwan, Kang Pil-Won, Chun Byung Won, Yoo Seong Yeon
Forensic DNA Division, National Forensic Service, Wonju, 26460, South Korea.
J Forensic Sci. 2020 Jan;65(1):209-213. doi: 10.1111/1556-4029.14147. Epub 2019 Aug 21.
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.
在商业多重短串联重复序列（STR）试剂盒中使用非FTA卡进行直接PCR时，建议使用特定缓冲液进行预处理步骤。在此，我们设计了一种利用非FTA卡（口腔细胞采样试剂盒）的快速直接PCR方法，省略了涉及Prep-n-Go™缓冲液的预处理步骤，并且该方法显示出与GlobalFiler™ Express PCR扩增试剂盒、GlobalFiler™ PCR扩增试剂盒和PowerPlex Fusion系统兼容。为了优化PCR条件，我们用不同的最终PCR体积和循环次数测试了该方法。最后，我们使用50份韩国颊拭子样本进行了性能测试，并证实了该方法的高性能，当在10 μL最终PCR体积中使用GlobalFiler™ PCR扩增试剂盒和PowerPlex Fusion系统在29个循环时，超过90%的样本能检测出完整图谱。因此，我们报告了一种简单的直接PCR设置，用于分析使用韩国制造的非FTA卡收集的参考样本。
