Inter-laboratory comparison of DNA preservation in archival paraffin-embedded human brain tissue from participating centres on four continents.
Author(s): Kösel S, Grasbon-Frodl EM, Arima K, Chimelli L, Hahn M, Hashizume Y, Hulette C, Ikeda K, Jacobsen PF, Jones M, Kobayashi M, Love S, Mizutani T, Rosemberg S, Sasaki A, Smith TW, Takahashi H, Vortmeyer AO, Graeber MB
Publication: Neurogenetics, 2001, Vol. 3, Page 163-70
PubMed ID: 11523568 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
-
Study Purpose
The purpose of this study was to determine the impact of different fixatives on the yield of DNA extracted from paraffin embedded brain specimens.
Summary of Findings:
The yield of DNA extracted from brain specimens preserved with buffered formalin or ethanol containing fixatives was significantly greater than specimens preserved with unbuffered formalin.
Biospecimens
Preservative Types
- Formalin
- Other Preservative
- Ethanol
Diagnoses:
- Alzheimer's Disease
- Parkinson's Disease
- Lewy Body Disease
- Down Syndrome
- Not specified
Platform:
Analyte Technology Platform DNA Fluorometry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Ethanol
Formalin (buffered)
Paraformaldehyde
Formalin (unbuffered)
-
Study Purpose
This study compared PCR efficacy of DNA extracted from differentially fixed brain specimens.
Summary of Findings:
PCR analysis of mitochondrial sequences was successful for 46% of differentially preserved brain specimens, and was significantly influenced by both the yield of extracted DNA and the type of fixative used, with specimens preserved in buffered formalin yielding the most robust results. Filtration of extracted DNA prior to PCR analysis improved both the success rate and band intensity. Further optimization led the authors to conclude that small DNA fragments (<300 bp) were inhibitory to the PCR reaction but could be attenuated by increasing template amount.
Biospecimens
Preservative Types
- Formalin
- Other Preservative
- Ethanol
Diagnoses:
- Alzheimer's Disease
- Parkinson's Disease
- Lewy Body Disease
- Down Syndrome
Platform:
Analyte Technology Platform DNA PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Ethanol
Formalin (buffered)
Paraformaldehyde
Formalin (unbuffered)
-
Study Purpose
The purpose of this study was to compare the quality of DNA extracted from formalin-fixed, paraffin-embedded and fresh frozen brain specimens via HPLC. One brain per preservation method was employed.
Summary of Findings:
HPLC analysis produced equivalent results for DNA extracted from fresh frozen and formalin-fixed paraffin-embedded specimens.
Biospecimens
Preservative Types
- Formalin
- Frozen
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform DNA HPLC Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
Snap frozen