Evaluation of the Effects of Different Sample Collection Strategies on DNA/RNA Co-Analysis of Forensic Stains

Genes (Basel). 2022 May 30;13(6):983. doi: 10.3390/genes13060983.

Abstract

The aim of this study was to evaluate the impact of different moistening agents (RNase-free water, absolute anhydrous ethanol, RNAlater®) applied to collection swabs on DNA/RNA retrieval and integrity for capillary electrophoresis applications (STR typing, cell type identification by mRNA profiling). Analyses were conducted on whole blood, luminol-treated diluted blood, saliva, semen, and mock skin stains. The effects of swab storage temperature and the time interval between sample collection and DNA/RNA extraction were also investigated. Water provided significantly higher DNA yields than ethanol in whole blood and semen samples, while ethanol and RNAlater® significantly outperformed water in skin samples, with full STR profiles obtained from over 98% of the skin samples collected with either ethanol or RNAlater®, compared to 71% of those collected with water. A significant difference in mRNA profiling success rates was observed in whole blood samples between swabs treated with either ethanol or RNAlater® (100%) and water (37.5%). Longer swab storage times before processing significantly affected mRNA profiling in saliva stains, with the success rate decreasing from 91.7% after 1 day of storage to 25% after 7 days. These results may contribute to the future development of optimal procedures for the collection of different types of biological traces.

Keywords: DNA/RNA co-extraction; STR typing; mRNA profiling; swab; trace collection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Coloring Agents* / analysis
  • DNA / analysis
  • DNA / genetics
  • Ethanol
  • RNA* / genetics
  • RNA, Messenger / genetics
  • Water

Substances

  • Coloring Agents
  • RNA, Messenger
  • Water
  • Ethanol
  • RNA
  • DNA

Grants and funding

This research was funded by Compagnia di San Paolo (funding “Progetti di Ricerca di Ateneo 2012”) and Fondazione CRT (grant no. 2013.2301 to C.R).