Simple salting-out method for DNA extraction from formalin-fixed, paraffin-embedded tissues
Author(s): Rivero Elena RC, Neves Adriana C, Silva-Valenzuela Maria G, Sousa Suzana OM, Nunes Fabio D
Publication: Pathol Res Pract, 2006, Vol. 202, Page 523
PubMed ID: 16723190 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
To assess DNA yield and size distribution after extraction from FFPE tissues using three distinct methods (sodium acetate, phenol-chloroform, and an unspecified commercial isolation kit).
Summary of Findings:
The quantity of DNA from FFPE tissue was comparable among extraction methods, with all methods producing adequate amounts of DNA. Further, the size distribution of DNA was identical among extraction methods.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA Electrophoresis DNA Spectrophotometry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Analyte Extraction and Purification Analyte isolation method Phenol-choloroform extraction
An unspecified commercial DNA isolation kit
Salting out extraction (ammonium acetate)
Simple salting out method (4M ammonium acetate)
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Study Purpose
To assess the molecular applicability of DNA extracted from FFPE tissue by different methods via PCR analysis of representative genes (beta-globin, adenomatous polyposi coli (APC)) and a spectrum of amplicon sizes (167-536 bp).
Summary of Findings:
PCR amplification of shorter products (<268 bp) was successful for all DNA extraction methods and cases examined, but amplification of longer products (536 bp) was successful on a case by case basis and did not appear to be method dependent.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Analyte Extraction and Purification Analyte isolation method Phenol-chloroform extraction
An unspecified commercial DNA isolation kit
Salting out extraction (ammonium acetate)
PCR Specific Targeted nucleic acid APC
beta-globin
PCR Specific Length of gene fragment 167 bp
268 bp
536 bp