Effect of bone decalcification procedures on DNA in situ hybridization and comparative genomic hybridization. EDTA is highly preferable to a routinely used acid decalcifier.
Author(s): Alers JC, Krijtenburg PJ, Vissers KJ, van Dekken H
Publication: J Histochem Cytochem, 1999, Vol. 47, Page 703-10
PubMed ID: 10219063 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to compare the effects of acid-based (RDO) and chelating-agent (EDTA) decalcification on ISH, CGH and flow cytometry results.
Summary of Findings:
While strong ISH signals were detected using centromereic probes on EDTA-decalcified specimens, RDO-decalcified specimens failed to show any signal. The authors report that there were too many nuclear debris to determine the DNA index by flow cytometry from RDO-decalcified specimens; however, EDTA-decalcified specimens showed distinct diploid and tetraploid peaks. DNA was not extracted from the RDO-treated bone, but fragments of up to 10 kB were extracted from the EDTA-decalcified specimens. Further, using EDTA-decalcified specimens, determination of loss and gain of sequences was possible by CGH.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Autopsy
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA Flow cytometry DNA In situ hybridization DNA CGH Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Analyte Extraction and Purification Decalcification solution/ duration Acid based agent (RDO)
Chelating agent (10% EDTA)