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Comparing Two automated Methods of dna extraction from Degraded Skeletal Remains

Forensic Medicine and Anatomy Research 2021年第2期9卷 24-30页

作者： Linda Rubinstein Independent Researcher Israel

dna extraction from degraded skeletal samples is often particularly challenging. The difficulty derives from the fact that variable environment has a significant effect on dna preservation. During the years 2002-2015 unidentified degraded skeletal remains were accumulated at our institute, National Institute of Forensic Medicine (NIFM), most of them with none or partial dna profile. As new methods rapidly emerge, we revisited these samples with partial dna profiles in the hope to add additional alleles and eventually be able to identify these previously unidentifiable samples. We have chosen to use these samples to compare two automated methods: Prepfiler Express BTA (Applied Biosystems) and QIAcube (Quiagen), in hope of acquiring a more complete dna profile and eventually make new identifications possibly comparing these profiles with missing person database. In both methods, a preparation step is required, after which the samples undergo automatic dna extraction. The two protocols are based on different extraction methods. Fresh or non-problematic bone samples as the positive control gave the same results in both methods. In the degraded skeletal samples, the results were significantly better using the QIAcube method in our hands, but since degraded samples are highly variable the combination of both methods could be useful to receive better and more reliable profiles.

关键词： automated dna extraction Degraded Skeletal Remains dna Profiling

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Improved resolution of mixed STR profiles using a fully automated differential cell lysis/dna extraction method

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Forensic Sciences Research 2020年第2期5卷 106-112页

作者： Matthew C.Goldstein Jordan O.Cox Lori B.Seman Tracey Dawson Cruz Department of Forensic Science Virginia Commonwealth UniversityRichmondVirginiaUSA

Sexual assault evidence often contains sperm cells,which are typically separated from nonsperm cells using manual differential lysis *** goal of this study was to evaluate the automated QIAGEN QIAcube for this purpose and to compare it to manual QIAGEN and manual organic differential methods using dna yields and STR profile data for *** yields were determined by qPCR,followed by multiplex STR amplification,CE analysis,and mixture *** automated method was capable of effective cell separation,producing dna yields sufficient for STR ***,sperm fraction human:male dna ratios from the QIAcube samples were consistently closer to the desired 1:1 and STR profiles were less likely to result in mixtures,with 6–8fewer female alleles detected(median 1.5 alleles).Ultimately,using the QIAcube for automated differential processing of semen-containing mixtures reduces the need for downstream mixture interpretation and improves STR profile quality with substantially less hands-on time.

关键词： Forensic sciences forensic genetics sexual assault evidence QIAcube differential lysis sperm lysis automated dna extraction mixture analysis

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