A high frequency of sequence alterations is due to formalin fixation of archival specimens.
Author(s): Williams C, Pontén F, Moberg C, Söderkvist P, Uhlén M, Pontén J, Sitbon G, Lundeberg J
Publication: Am J Pathol, 1999, Vol. 155, Page 1467
PubMed ID: 10550302 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
To determine if PCR amplification efficiency is reduced when using DNA extracted from FFPE tissue in comparison to fresh frozen tissue.
Summary of Findings:
The number of microdissected cells from frozen tissue (10 to 200 cells per PCR reaction) did not affect successful PCR amplification, although amplification of longer products (350 bp) was unsuccessful with lower quantities of cells (10-20 cells per PCR reaction) microdissected from FFPE tissue.
Biospecimens
Preservative Types
- Formalin
- Frozen
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Snap frozen
Formalin (buffered)
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Study Purpose
To determine if the incidence of nonreproducible mutations is dependent upon tissue fixation method (FFPE versus fresh frozen tissue).
Summary of Findings:
Fixative-dependent effects were observed with DNA sequence analysis. While two known mutations were detected in frozen tissue, DNA extracted from FFPE tissue produced a total of 28 nonreproducible mutations that average 1 per 500 cells. These artifactual mutations included silent (28%) and missense or nonsense (72%) mutations.
Biospecimens
Preservative Types
- Formalin
- Frozen
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA DNA sequencing Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
Snap frozen