Validation of EZ1® Advanced XL workstation using EZ1 Investigator Kit

Autor: Abramović, Marko Krešimir, Barbarić, Lucija, Valić, Marija, Merkaš, Siniša, Rožić, Sara, Sukser, Viktorija, Uvodić, Petra, Crnjac, Josip, Popović, Maja, Ortynski, Nataša, Mršić, Gordan
Přispěvatelé: Kayser M., Ordog T., Vuk-Pavlović S., Primorac D., Schanfield M.
Jazyk: angličtina
Rok vydání: 2015
Předmět:
Popis: With the ongoing need to constantly improve techniques that laboratories use for isolation and purification of DNA, we introduce EZ1® Advanced XL instrument for automated isolation and purification of DNA alongside with EZ1 Investigator Kit into our laboratory casework. The aim of this study was to compare existing Chelex®100 method with EZ1® Advanced XL workstation, and to determine accuracy, reliability, and flexibility of EZ1 Investigator kit. Genomic DNA was isolated from whole blood, buccal swabs and forensic samples. The genomic DNA content of each sample was determined by quantitative real-time polymerase chain reaction (PCR) using Quantifiler® Human DNA Quantification Kit and Investigator Quantiplex kit, respectively. Amplification was performed using AmpFlSTR®NGMTM PCR Amplification Kit. Results were presented as value of DNA concentrations, mean value of peak heights, mean value of ratio of peak heights and percentage of samples that gave full DNA profile, following standard forensic eligibility criteria. All samples showed average ratio of peak heights for heterozygous loci above 60%, which indicates good DNA profile quality. Isolation, amplification and detection of DNA from cigarette butts and chewing gums (forensic samples) gave better results when using EZ1 workstation. Samples of whole blood showed equally good results using Chelex®100 and EZ1 workstation. Only samples of buccal swabs gave better results with Chelex method. Apart from this, and although future studies with different kinds of samples are needed, all criteria were met and EZ1® Advanced XL instrument with EZ1 Investigator Kit is introduced into our every day casework.
Databáze: OpenAIRE