Comparative evaluation of different extraction and quantification methods for forensic RNA analysis |
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| Affiliation: | 1. Institute of Legal Medicine, University of Bonn, Stiftsplatz 12, D-53111 Bonn, Germany;1. Forensic Research and Development, Institute of Environmental Science and Research Limited, Auckland, New Zealand;2. New Zealand Genomics Ltd./Bioinformatics Institute, School of Biological Sciences, University of Auckland, Auckland, New Zealand;1. Institute of Forensic Medicine, University of Zurich, Zurich, Switzerland;2. National Center for Forensic Science, University of Central Florida, Orlando, USA;3. Department of Chemistry, University of Central Florida, Orlando, USA;1. Menarini Silicon Biosystems S.p.A., Bologna, Italy;2. Reparto Investigazioni Scientifiche Carabinieri R.I.S., Roma, Italy;1. Institute for Forensic Medicine, Sichuan University, Chengdu, Sichuan, China;2. Department of Criminal Science and Technology, Sichuan Police College, Luzhou, Sichuan, China |
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| Abstract: | Since about 2005, there is increasing interest in forensic RNA analysis whose versatility may very favorably complement traditional DNA profiling in forensic casework. There is, however, no method available specifically dedicated for extraction of RNA from forensically relevant sample material.In this study we compared five commercially available and commonly used RNA extraction kits and methods (mirVana™ miRNA Isolation Kit Ambion; Trizol® Reagent, Invitrogen; NucleoSpin® miRNA Kit Macherey-Nagel; AllPrep DNA/RNA Mini Kit and RNeasy® Mini Kit both Qiagen) to assess their relative effectiveness of yielding RNA of good quality and their compatibility with co-extraction of DNA amenable to STR profiling.We set up samples of small amounts of dried blood, liquid saliva, semen and buccal mucosa that were aged for different time intervals for co-extraction of RNA and DNA. RNA quality was assessed by determination of ‘RNA integrity number’ (RIN) and quantitative PCR based expression analysis. DNA quality was assessed via monitoring STR typing success rates.By comparison, the different methods exhibited considerable differences between RNA and DNA yields, RNA quality values and expression levels, and STR profiling success, with the AllPrep DNA/RNA Mini Kit and the NucleoSpin® miRNA Kit excelling at DNA co-extraction and RNA results, respectively.Overall, there was no ‘best’ method to satisfy all demands of comprehensible co-analysis of RNA and DNA and it appears that each method has specific merits and flaws. We recommend to cautiously choose from available methods and align its characteristics with the needs of the experimental setting at hand. |
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| Keywords: | RNA extraction RNA quantification RNA/DNA co-extraction Forensic RNA analysis |
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