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Transcriptomic analysis of degraded forensic body fluids.
Meng-Han Lin1, Daniel F Jones2, Rachel Fleming1
1Forensic Research and Development, Institute of Environmental Science and Research Limited, Auckland, New Zealand.
Forensic Science International. Genetics
|March 24, 2015
Summary
Massively parallel sequencing effectively analyzes degraded RNA from forensic samples like saliva and blood. This technology enables accurate body fluid identification even from aged, low-abundance biological evidence.
Area of Science:
- Forensic science
- Molecular biology
- Genomics
Background:
- Degraded RNA analysis is challenging in forensic science.
- Forensic samples are often degraded and low in quantity.
- RNA analysis can identify body fluids via specific transcripts.
Purpose of the Study:
- To demonstrate Massively Parallel Sequencing (MPS) for degraded RNA analysis.
- To assess MPS performance on forensically relevant body fluids.
- To characterize RNA degradation patterns in different body fluids over time.
Main Methods:
- RNA extraction from fresh, 2-week, and 6-week aged samples of oral mucosa/saliva, circulatory blood, menstrual blood, and vaginal fluid.
- Massively Parallel Sequencing (MPS) of extracted RNA.
- Bioinformatic analysis including read alignment to the human genome and Fragments Per Kilobase of exon per Million fragments mapped (FPKM) calculation.
Main Results:
- High-quality sequencing output (>80% sequence above Q30) was achieved despite significant RNA degradation.
- Over 80% of reads successfully aligned to the human reference genome for most samples.
- FPKMs confirmed accurate detection of body fluid-specific markers.
- Differential RNA degradation patterns were observed across different body fluid types over time.
Conclusions:
- MPS is a practical and effective technology for analyzing degraded RNA from minimal forensic samples.
- The study validates MPS for accurate body fluid identification in forensic investigations.
- Understanding RNA degradation profiles aids in interpreting results from aged forensic samples.
