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

• Record Type: Journal Article
• Authors: Jung, Ju Yeon; Ryu, Ga Hee; Kim, Da‐Hye; Kim, Joo‐Young; Hwang, In Kwan; Kang, Pil‐Won; Chun, Byung Won; Yoo, Seong Yeon
• Is Part Of: Journal of forensic sciences. Volume 65:Issue 1(2020)
• Page Start: 209
• Page End: 213
• Publication Date: 2019-08-21
• Subjects: forensic science -- DNA typing -- direct polymerase chain reaction -- Oral Cell Sampling Kit -- Prep‐n‐Go™ Buffer -- reference samples; Medical jurisprudence -- Periodicals; Forensic sciences -- Periodicals; Forensic Medicine -- Periodicals; Gerechtelijke geneeskunde; Gerechtelijke chemie; Gerechtelijke psychiatrie; 363.2505
• DOI: 10.1111/1556-4029.14147
• Languages: English
• ISSNs: 0022-1198

Abstract

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.