Feasibility of archival non-buffered formalin-fixed and paraffin-embedded tissues for PCR amplification: an analysis of resected gastric carcinoma.
Author(s): Inoue T, Nabeshima K, Kataoka H, Koono M
Publication: Pathol Int, 1996, Vol. 46, Page 997-1004
PubMed ID: 9110353 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine the effects of fixation duration, storage, and gene fragment length on PCR success in gastric carcinomas.
Summary of Findings:
Amplification of a 190 bp fragment of p53 was successful in 6 of 6 specimens fixed for 1 day, 7 of 16 specimens fixed for 2-3 days, 4 of 18 specimens fixed for 4-6 days, and 0 of 34 specimens fixed for more than 7 days. Storage of paraffin blocks did not affect specimens fixed for 1 day, but in specimens fixed for 2 days or more, storage for more than 30 months reduced PCR success. Electrophoresis revealed that the peak size of extracted DNA decreased from 265 bp in specimens fixed for 1 day to 180 bp in specimens fixed for 10 days. When amplification of three different size fragments of Beta globin was performed on specimens fixed for 1 day, product decreased with increasing target size in one specimen, but in the other specimen all products were produced equally.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA Electrophoresis DNA PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Time in fixative 1 day
2 days
3 days
4 days
5 days
6 days
7 days
8 days
9 days
10 days
PCR Specific Targeted nucleic acid p53
Beta globin
PCR Specific Length of gene fragment 167 bp
190 bp
262 bp
345 bp
Storage Storage duration <30 months
>30 months
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Study Purpose
The purpose of this study was to determine the effects formic acid build-up in unbuffered formalin on PCR success rates in liver specimens.
Summary of Findings:
PCR success rates decreased in specimens fixed for 6 h in unbuffered formalin containing increasing concentrations of added formic acid compared to specimens fixed in fresh unbuffered formalin without formic acid.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Autopsy
- Not specified
Platform:
Analyte Technology Platform DNA PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Fixative additive/buffer Formic acid
Multiple concentrations evaluated