A Comparison of DNA Extraction Using AutoMate Express™ and EZ1 Advanced XL from Liquid Blood, Bloodstains, and Semen Stains
Corresponding Author
Koji Fujii Ph.D.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Additional information and reprint requests:
Koji Fujii, Ph.D.
Senior Researcher
Fourth Biological Section, National Research Institute of Police Science
6-3-1 Kashiwanoha, Kashiwa
Chiba 277-0882
Japan
E-mail: [email protected]
Search for more papers by this authorShota Inokuchi B.S.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorTetsushi Kitayama Ph.D.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorHiroaki Nakahara Ph.D.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorNatsuko Mizuno Ph.D.
Fifth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorKazumasa Sekiguchi Ph.D.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorCorresponding Author
Koji Fujii Ph.D.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Additional information and reprint requests:
Koji Fujii, Ph.D.
Senior Researcher
Fourth Biological Section, National Research Institute of Police Science
6-3-1 Kashiwanoha, Kashiwa
Chiba 277-0882
Japan
E-mail: [email protected]
Search for more papers by this authorShota Inokuchi B.S.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorTetsushi Kitayama Ph.D.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorHiroaki Nakahara Ph.D.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorNatsuko Mizuno Ph.D.
Fifth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorKazumasa Sekiguchi Ph.D.
Fourth Biological Section, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba 277-0882, Japan
Search for more papers by this authorAbstract
In this study, DNA was extracted using an AutoMate Express™ and an EZ1 Advanced XL from liquid blood, fresh and aged bloodstains, and fresh and aged semen stains. Extracted DNA was quantified by real-time PCR using the D17Z1 locus. Short tandem repeat typing was performed using an AmpFℓSTR® Identifiler kit. The yields of DNA obtained by the AutoMate Express™ were higher from fresh bloodstains and fresh semen stains, almost the same from aged bloodstains and aged semen stains, but slightly lower from liquid blood compared with those obtained by the EZ1 Advanced XL. The addition of dithiothreitol or the use of PrepFiler™ lysis buffer improved the EZ1 Advanced XL results from fresh bloodstains, but not for liquid blood and aged bloodstains. Our results demonstrated that the PrepFiler™ lysis buffer is the main contributor to the higher DNA yields of the AutoMate Express™ for fresh bloodstains.
References
- 1Scherczinger CA, Bourke MT, Ladd C, Lee HC. DNA extraction from liquid blood using QIAamp. J Forensic Sci 1997; 42: 893–6.
- 2Greenspoon SA, Ban JD, Sykes K, Ballard EJ, Edler SS, Baisden M, et al. Application of the BioMek 2000 laboratory automation workstation and the DNA IQ system to the extraction of forensic casework samples. J Forensic Sci 2004; 49: 29–39.
- 3Anslinger K, Bayer B, Rolf B, Keil W, Eisenmenger W. Application of the BioRobot EZ1 in a forensic laboratory. Leg Med 2005; 7: 164–8.
- 4Montpetit SA, Fitch IT, O'Donnell PT. A simple automated instrument for DNA extraction in forensic casework. J Forensic Sci 2005; 50: 555–63.
- 5Kishore R, Reef HW, Anderson VJ, Sanchez NA, Buoncristiani MR. Optimization of DNA extraction from low-yield and degraded samples using the BioRobot EZ1 and BioRobot M48. J Forensic Sci 2006; 51: 1055–61.
- 6Krnajski Z, Geering S, Steadman S. Performance verification of the Maxwell 16 instrument and DNA IQ reference sample kit for automated DNA extraction of known reference samples. Forensic Sci Med Pathol 2007; 3: 264–9.
- 7Frégeau CJ, Lett CM, Elliott J, Yensen C, Fourney RM. Automated processing of forensic casework samples using robotic workstations equipped with nondisposable tips: contamination prevention. J Forensic Sci 2008; 53: 632–51.
- 8Hoffman E, Ringberg M. Feasibility study to develop a method for automating differential extraction and DNA isolation: a comparison of automated systems. Proceedings of the Twenty First International Symposium on Human Identification; 2010 Oct 11–14; San Antonio, TX, 2010, http://www.promega.ca/products/pm/genetic-identity/ishi-conference-proceedings/proceedings-index-home/?origUrl=http%3a%2f%2fwww.promega.com%2fgeneticidproc%2f (accessed December 16, 2011).
- 9Turnbough MA, Davis CP, King JL, Dixon B, Liu JY, Budowle B, et al. Competitive extraction robot audit: a comparison of the Applied Biosystems™ AutoMate Express™, QIAgen® EZ1® Advanced XL, and the Promega Maxwell® 16. Proceedings of the Twenty First International Symposium on Human Identification; 2010 Oct 11–14; San Antonio, TX, 2010, http://www.promega.ca/products/pm/genetic-identity/ishi-conference-proceedings/proceedings-index-home/?origUrl=http%3a%2f%2fwww.promega.com%2fgeneticidproc%2f (accessed December 16, 2011).
- 10Bogas V, Balsa F, Carvalho M, Anjos MJ, Pinheiro MF, Corte-Real F. Comparison of four DNA extraction methods for forensic application. Forensic Sci Int-Gen Supplement Series 2011; 3: e194–5.
10.1016/j.fsigss.2011.08.097 Google Scholar
- 11Davis CP, King JL, Budowle B, Eisenberg AJ, Turnbough MA. Extraction platform evaluations: a comparison of Automate Express™, EZ1® Advanced XL, and Maxwell® 16 bench-top DNA extraction systems. Leg Med 2012; 14: 36–9.
- 12Brevnov MG, Pawar HS, Mundt J, Calandro LM, Furtado MR, Shewale JG. Developmental validation of the PrepFiler™ Forensic DNA extraction kit for extraction of genomic DNA from biological samples. J Forensic Sci 2009; 54: 599–607.
- 13 Applied Biosystems. Product bulletin: PrepFiler™ Forensic DNA extraction kit. Foster City, CA: Applied Biosystems, 2008, http://www3.appliedbiosystems.com/cms/groups/applied_markets_marketing/documents/generaldocuments/cms_057536.pdf (accessed June 21, 2012).
- 14Fujii K, Inokuchi S, Kitayama T, Nakahara H, Mizuno N, Sekiguchi K. DNA extraction from various forensic samples using automated extraction instruments. Jpn J Forensic Sci Tech 2012; 17: 15–26.
10.3408/jafst.17.15 Google Scholar
- 15 Applied Biosystems. PrepFiler™ Express™ and PrepFiler™ Express BTA™ Forensic DNA extraction kits user guide (rev.C). Foster City, CA: Applied Biosystems, 2012, http://www3.appliedbiosystems.com/cms/groups/applied_markets_support/documents/generaldocuments/cms_085891.pdf (accessed June 21, 2012).
- 16 Applied Biosystems. PrepFiler™ Forensic DNA extraction kit user guide (rev.B). Foster City, CA: Applied Biosystems, 2008, http://www3.appliedbiosystems.com/cms/groups/applied_markets_support/documents/generaldocuments/cms_053966.pdf (accessed June 21, 2012).
- 17 QIAgen. EZ1® DNA investigator handbook, 4th edn. Hilden, Germany: QIAgen, 2009, http:www.qiagen.com (accessed June 21, 2012).
- 18Nakahara H, Fujii K, Mizuno N, Yoshida K, Kasai K. Evaluations of DNA quantification methods for forensic biological samples. Jpn J Forensic Sci Tech 2007; 12: 13–26.
10.3408/jafst.12.13 Google Scholar
- 19Yoshida K, Sekiguchi K, Kasai K, Sato H, Seta S, Sensabaugh GF. Evaluation of new primers for CSF1PO. Int J Legal Med 1997; 110: 36–8.
- 20Fujii K, Senju H, Yoshida K, Sato H, Kasai K. A new procedure for the multiplex PCR amplification of TH01, CSF1PO and TPOX loci with small amplicon lengths. Jpn J Sci Tech Iden 2002; 6: 129–34.
10.3408/jasti.6.129 Google Scholar
- 21Butler JM, Shen Y, McCord BR. The development of reduced size STR amplicons as tools for analysis of degraded DNA. J Forensic Sci 2003; 48: 1054–64.
- 22Sekiguchi K, Imaizumi K, Matsuda H, Mizuno N, Yoshida K, Senju H, et al. MtDNA sequence analysis using capillary electrophoresis and its application to the analysis of MtDNA in hair. Jpn J Sci Tech Iden 2003; 7: 123–30.
10.3408/jasti.7.123 Google Scholar
- 23Matsuda H, Imaizumi K, Kubota S, Miyasaka S, Yoshino M, Seta S. Technical investigation of DNA extraction from single hair shaft. Reports of the National Research Institute of Police Science 1997; 50: 23–8.
- 24 Applied Biosystems. Product bulletin: AutoMate Express™ Forensic DNA extraction system. Foster City, CA: Applied Biosystems, 2010, http://www3.appliedbiosystems.com/cms/groups/applied_markets_marketing/documents/generaldocuments/cms_086180.pdf (accessed June 25, 2012).
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