Automation of DNA and miRNA co-extraction for miRNA-based identification of human body fluids and tissues

In the last years, microRNA (miRNA) analysis came into focus in the field of forensic genetics. Yet, no standardized and recommendable protocols for co‐isolation of miRNA and DNA from forensic relevant samples have been developed so far. Hence, this study evaluated the performance of an automated Ma...

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Bibliographic Details
Published in:Electrophoresis Vol. 37; no. 21; pp. 2742 - 2750
Main Authors: Kulstein, Galina, Marienfeld, Ralf, Miltner, Erich, Wiegand, Peter
Format: Journal Article
Language:English
Published: Germany Blackwell Publishing Ltd 01-10-2016
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Summary:In the last years, microRNA (miRNA) analysis came into focus in the field of forensic genetics. Yet, no standardized and recommendable protocols for co‐isolation of miRNA and DNA from forensic relevant samples have been developed so far. Hence, this study evaluated the performance of an automated Maxwell® 16 System‐based strategy (Promega) for co‐extraction of DNA and miRNA from forensically relevant (blood and saliva) samples compared to (semi‐)manual extraction methods. Three procedures were compared on the basis of recovered quantity of DNA and miRNA (as determined by real‐time PCR and Bioanalyzer), miRNA profiling (shown by Cq values and extraction efficiency), STR profiles, duration, contamination risk and handling. All in all, the results highlight that the automated co‐extraction procedure yielded the highest miRNA and DNA amounts from saliva and blood samples compared to both (semi‐)manual protocols. Also, for aged and genuine samples of forensically relevant traces the miRNA and DNA yields were sufficient for subsequent downstream analysis. Furthermore, the strategy allows miRNA extraction only in cases where it is relevant to obtain additional information about the sample type. Besides, this system enables flexible sample throughput and labor‐saving sample processing with reduced risk of cross‐contamination.
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Supplementary Table 1: Overview of genuine case work samples. Supplementary Table 2: Overall evaluation of procedures. Supplementary Figure 1: DNA and miRNA yields in pg/μL of aged stains. Dried blood spots were obtained from blood cards of corpses from 2006. Five cards were analyzed by automated co-extraction of miRNA and DNA.
See the article online to view Figs. 2–4 in colour.
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ISSN:0173-0835
1522-2683
DOI:10.1002/elps.201600365