PowerPlex^® 16 HS System의 유효성 확인
Internal Validation of the PowerPlex 16 HS System
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참고문헌
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초록
본 연구는 PowerPlex^® 16 HS System (Promega)의 감정적용에 앞서 법의유전학 분야의 사건증 거물들을 분석할 때 적절한 수준의 실험결과를 얻을 수 있는지 확인하는데 그 목표를 두고 있다. 구강상피 세포와 혈액 시료에서 PrepFilerô Forensic DNA Extraction kit로 추출한 DNA를 Quantifiler Duoô Human DNA Quantification kit와 7500 real time PCR system을 이용하여 정량하였다. 정량된 DNA로써 유효성 검증을 위한 민감성, 재현성, 정확성, 혼합반 및 오염에 대한 실험에 사용하였다. 유전자 증폭은 9700 GeneAmp PCR system을 이용하였고, 유전자형 분석은 3130 Genetic Analyzer로 하였다. 그 결과, 1.0 ng DNA로 시행한 반복실험에서 0.07 bp 이하의 표준편차를 보였고, 62.5 pg이상의 template DNA에서 전체 유전자형이 분석되었다. 혼합반의 경우, 1:9의 90%이상에서 부공여자의 유전자형이 분석되었고 유전자형이 알려진 대조 시료와 검증되지 않은 증거물 시료의 유전자형을 비교분석한 결과, 서로 일치함을 확인할 수 있었다.
Abstract
The aim of this study is to verify the consistency of the results and optimize the conditions in our lab. DNA samples extracted from a buccal swab and blood by using the PrepFilerTM DNA extraction kit were prepared for quantification by the Tecan Freedom EVO automated liquid handler. The extracts were measured by real time PCR performed by the Quantifiler DuoTM‚ Human DNA Quantification Kit and 7500 real time PCR system. Internal validations on reproducibility, precision, sensitivity, mixture and contamination were performed according to the revised SWGDAM guidelines. After four runs of 1.0 ng DNA templates,all alleles showed standard deviations of less than 0.07 bp. Full profiles were successfully analyzed in all runs over 0.0625 ng. Full profiles of minor contributors were detected more than 90% at 1:9 and over mixtures. The comparison between known reference and non-probative evidence samples showed perfect overall concordance.
참고문헌 (13)
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