Highlights
- •Comparison of five different methods for forensic RNA/DNA co-extraction.
- •No method found optimal for all aspects of RNA/DNA co-analysis.
- •Best RNA-results: NucleoSpin® miRNA Kit.
- •Best DNA-results: AllPrep DNA/RNA Mini Kit.
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.
Keywords
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Article info
Publication history
Published online: January 16, 2015
Accepted:
January 15,
2015
Received in revised form:
November 20,
2014
Received:
September 5,
2014
Identification
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© 2015 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
