A Comparison of DNA Extraction Using AutoMate Express™ and EZ1 Advanced XL from Liquid Blood, Bloodstains, and Semen Stains

Abstract

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.

Comparison of an automated nucleic acid extraction system with the column-based procedure

“…Successful EZ1-based DNA extraction from formalin-fi xed, paraffi nembedded tissue samples [9], or otherwise long-term preserved tissue specimens [8] has been demonstrated. Even DNA extraction from previously stained smears with the EZ1 system works reliably [6]. The EZ1 system extracts not only DNA but also RNA depending on the chosen program [1].…”

D5S818 Typing Discrepancy Between PowerPlex® Fusion and Other STR Kits Including GlobalFiler® Caused by a One‐base Deletion in 31 Nucleotides Upstream of the Repeat Region

“…The stored DNA of the Sample A was used in this study. This DNA sample was extracted from liquid blood using a MagNA Pure LC instrument (Roche Diagnostics, Basel, Switzerland) for the population studies and its 1/10 solution diluted with low TE buffer (TE −4 ) was quantified in real‐time PCR assay using the 207‐bp amplicon at D17Z1 locus in the same method described previously in the comparison study of DNA extraction instruments . The sample was used with the approval of the ethics committee at our institute.…”

Integrated Polymerase Chain Reaction Technologies (Sample-to-Answer Technologies)

“…Regardless of the type of specimen being analyzed, however, the first step in any PCR-based molecular genetic assay was the preparation of a nucleic acid extract. While this was originally performed manually (typically by boiling or chemical lysis, that is, the use of noxious organic solvents like phenolechloroform and ethanol precipitation), a wide variety of commercially available instruments have since been developed that automate this process (Fujii et al, 2013). Automated cell lysis can be achieved through the use of detergents and chaotropic salts, heat, mechanical disruption, or simple pressure.…”