PrepFiler™ Forensic DNA Extraction Kit를 이용한 DNA 추출 시험방법의 유효화
Internal Validation of Manual and Automated DNA Extraction Method from Using the PrepFiler™ Forensic DNA Extraction Kit
1
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
본 연구에서는 상용화 DNA 추출 kit 중 yield와 회수율이 높은 것으로 알려진 PrepFilerTM Forensic DNA Extraction Kit (Applied Biosystems, Foster City, CA)을 이용한 DNA 추출 방법의 시험방법 유효화 연구를 진행하였다. DNA 추출방법은 수동 방법과 자동화 방법을 비교하여 진행하였다. DNA 추출 방법의 비교를 위해 사용된 시료는 전혈 혈액(whole blood)과 타액시료(Liquid saliva)이다. 두 시험 방법에 따라 전혈 혈액의 Human DNA와 Male DNA 정량 값을 비교한 결과 0.1 μL에서 5 μL까지는 수동 유전자 추출 시험방법과 자동화 유전자추출 시험방법 간에 차이가 거의 없이 선형으로 증가하는 것으로 나타났으며, 10 μL에서는 수동 방법이 계속 선형으로 증가 했지만 자동화 방법은 감소하는 것으로 나타났다. 타액시료의 Human DNA와 Male DNA 정량 값을 비교한 결과 0.1 μL에서 10 μL까지는 수동 방법과 자동화 방법 간에 차이가 거의 없이 선형으로 증가하는 것으로 나타났다.
Abstract
In order to introduce a high yield and recovery DNA extraction method into our laboratory, a validation study of using the PrepFilerTM‚ Forensic DNA Extraction Kit (Applied Biosystems, Foster City, CA) was performed. We have validated protocols for the DNA extraction method used in manual and automated protocols. In the DNA extraction method, the samples that were compared were whole blood and liquid saliva. According to both methods, quantitative results using up to 0.1-5 μL human DNA from a sample of whole blood and male DNA showed no difference of a linear increase in the manual and automated method. However, there was a decrease for automated method for 10 μL. Also a comparison of quantitative results using a liquid saliva sample of human DNA and male DNA for up to 0.1-10 μL showed no difference of linear increase in the manual and automated method.
참고문헌 (12)
- J A Thomsona, "Validation of short tandem repeat analysis for the investigation of cases of disputed paternity", Forensic Sci Int, 100, pp.1~16 (1999)
- F Stanciu, "Automated DNA isolation for databasing purposes", Rom J Leg Med 17, 1, pp.51~58 (2009)
- A Barbaro, "Validation of Prep- FilerTM‚ forensic DNA extraction kit (Applied Biosystems)", Forensic Sci Int: Genetics Supplement Series, 2, pp.176~177 (2009)
- M G Brevnov, "Developmental Validation of the PrepFilerTM‚ Forensic DNA Extraction Kit for Extraction of Genomic DNA from Biological Samples", J Forensic Sci, 54(3), pp.599~607 (2009)
- "Revised Validation Guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM). Forensic Sci Commun 2004;6(3)"[URL]
- Applied Biosystems, "PrepFilerTM‚ Forensic DNA Extraction Kit User Guide. Part No. 4390932 Rev. B", ~ (2008)
- Applied Biosystems, "PrepFilerTM‚ Automated Forensic DNA Extraction Kit getting started guide. PPN 4393917 Rev. B", ~ (2008)
- M Brevnov, "Automated Extraction of DNA from Forensic Sample Types Using the PrepFilerTM‚ Automated Forensic DNA Extraction Kit", JALA, 10, pp.294~302 (2009)
- P Zimmermann, "Adaptation and evaluation of the PrepFilerTM‚ DNA extraction technology in an automated forensic DNA analysis process with emphasis on DNA yield, inhibitor removal and contamination security", Forensic Sci Int: Genetics Supplement Series, 2, pp.62~63 (2009)
- S A Greenspoon, "Automated PCR setup for forensic casework samples using the Normalization Wizard and PCR Setup robotic methods", Forensic Sci Int, 164, pp.240~248 (2006)
- M W Perlin, "Automated STR Data Analysis: Validation Studies", Cybergenetics, 1, pp.1~24 (2001)
- H Niederstatter, "A modular real-time PCR concept for determining the quantity and quality of human nuclear and mitochondrial DNA", Forensic Sci Int Genet, 1, pp.29~34 (2007)