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DNA extraction and quantification from touch and scrape preparations obtained from autopsy liver cells
C N M Ribeiro1, L C Peres, J M Pina-Neto
1Departamento de Patologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirao Preto, SP, Brazil.
Summary
This study presents a simple, low-cost method for collecting and fixing pediatric autopsy cells, yielding sufficient DNA for genetic analysis. 95% ethanol fixation of scrape preparations proved most effective for DNA extraction.
Area of Science:
- Forensic Science
- Molecular Biology
- Genetics
Background:
- Accurate DNA extraction from pediatric autopsy samples is crucial for genetic studies.
- Traditional methods for sample preservation can be costly and labor-intensive.
Purpose of the Study:
- To develop a simplified, cost-effective method for collecting and fixing pediatric autopsy cells.
- To assess the quantitative and qualitative adequacy of DNA extracted using this method.
Main Methods:
- Touch and scrape preparations of pediatric liver cells from 15 autopsies were fixed using 95% ethanol or methanol:acetic acid.
- DNA was extracted using the Wizard genomic DNA purification kit and quantified.
- Multiplex PCR was used to amplify azoospermia factor genes from a subset of samples.
Main Results:
- DNA yield ranged from 20 to 8,640 µg, with 95% ethanol fixation of scrape preparations yielding the most DNA.
- Extracted DNA quantities exceeded requirements for PCR (50 ng) and Southern blot (500 ng).
- No qualitative DNA differences were observed between fixatives or storage durations up to 24 months.
Conclusions:
- Touch and scrape preparations fixed in 95% ethanol provide a practical, low-cost, and reliable source of DNA from pediatric autopsy samples.
- This method offers advantages over cell culture, paraffin embedding, or freezing, requiring less specialized equipment and expertise.
- The method is suitable for long-term storage and genetic analyses, including PCR and Southern blot.