PCR analysis in archival postmortem tissues.
Author(s): Bonin S, Petrera F, Niccolini B, Stanta G
Publication: Mol Pathol, 2003, Vol. 56, Page 184
PubMed ID: 12782767 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The authors compared amplification results for ApoE and two fragment lengths of TTR (TTR1 = 291 and TTR4 = 339 base pairs) with and without several pre-PCR DNA treatment steps in three biopsy and nine autopsy biospecimens.
Summary of Findings:
No amplification was obtained for ApoE in any postmortem biospecimens regardless of treatment. TTR amplification was successful only in restored and denatured postmortem biospecimens: the 291bp fragment in 6 out of 9, and the 339 fragment in 1 out of 9. A pre-PCR DNA ligation step did not alter amplification results. ApoE and both TTR fragments were successfully amplified in all biopsy biospecimens regardless of treatment. The authors note that the underlying principal of the restoration steps outlined is the repair of DNA degradation-induced single strand breaks using PCR.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Autopsy
- Not specified
Platform:
Analyte Technology Platform DNA PCR DNA Spectrophotometry DNA Electrophoresis Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Analyte Extraction and Purification Analyte isolation method With a pre-PCR restoration treatment
Without a pre-PCR restoration treatment
With a DNA denaturation step
Without a DNA denaturation step
With a DNA ligation step
Without a DNA ligation step
Analyte Extraction and Purification Analyte isolation method With a DNA denaturation step
Without a DNA denaturation step
Analyte Extraction and Purification Analyte isolation method With a DNA ligation step
Without a DNA ligation step
PCR Specific Length of gene fragment 291 bp
339 bp
PCR Specific Targeted nucleic acid Apolipoprotein E
Prealbumin gene (TTR)
Biospecimen Acquisition Method of tissue acquisition Autopsy
Biopsy