Development of a Method of Suspension RNA-FISH for Forensically Relevant Epithelial Cells Using LNA Probes

• NCJ Number: 245539
• Journal: Forensic Science International: Genetics Volume: 9 Dated: March 2014 Pages: 85-92
• Author(s): Eletra Williams; Meng-Han Lin; SallyAnn Harbison; Rachel Fleming
• Date Published: March 2014
• Length: 8 pages
• Sponsoring Agency: Institute of Environmental Science and Research Ltd.
• Publication Format: Article
• Publication Type: Research (Applied/Empirical); Report (Study/Research)
• Language: English
• Country: Ireland

Annotation

This study developed and optimized a method that uses a locked nucleic acid (LNA) probe for the keratin 10 (KRT 10) mRNA for the labeling of vaginal and buccal epithelial cells in fresh and aged samples of mixed cells.

Abstract

The proposed method involves RNA suspension-fluorescent in situ hybridization (RNA S-FISH), using a LNA probe for the KRT 10, which is suitable as a potential marker for epithelial cells. Testing results show that RNA S-FISH labeling is compatible with subsequent DNA profiling. This was demonstrated by obtaining full DNA profiles from fresh RNA S-FISH labeled cell samples and partial DNA profiles from aged cell samples after LMD. This method provides flexibility in allowing for the analysis of a range of sample types and ages. In addition, it can be modified to minimize cell loss, maximize signal detection, or increase specificity. The detailed discussion of methods addresses cell sampling, recovery, and fixation, FISH probes and hybridization conditions, microscopy, controls, and DNA analysis and LMD. 1 table, 5 figures, and 60 references