PHOTOPROBER® Biotin: An Alternative Method for Labeling Archival DNA for Comparative Genomic Hybridization
| Autor: | Ulrike Bockmühl, Iver Petersen, Dirk Korinth, Konrad Donhuijsen |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2004 |
| Předmět: |
Cancer Research
Cytoplasm Light Biotin In situ hybridization Biology Genome lcsh:RC254-282 Pathology and Forensic Medicine chemistry.chemical_compound Nucleic acid thermodynamics Cell Line Tumor Neoplasms Image Processing Computer-Assisted FFPE tissue Humans Nick translation lcsh:QH573-671 In Situ Hybridization CGH Chromosome Aberrations Electrophoresis Agar Gel lcsh:Cytology Hybridization probe Nucleic Acid Hybridization Cell Biology General Medicine DNA DNA Neoplasm lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens Molecular biology non‐enzymatic labeling chemistry Biotinylation Karyotyping Molecular Medicine Other DNA Probes Comparative genomic hybridization |
| Zdroj: | Cellular Oncology : the Official Journal of the International Society for Cellular Oncology Cellular Oncology, Vol 26, Iss 5-6, Pp 329-334 (2004) |
| ISSN: | 1875-8606 1570-5870 |
| Popis: | Comparative genomic hybridization (CGH) represents a powerful method for screening the entire genome of solid tumors for chromosomal imbalances. Particularly it enabled the molecular cytogenetic analysis of archival, formalin‐fixed, paraffin‐embedded (FFPE) tissue. A well‐known dilemma, however, is the poor DNA quality of this material with fragment sizes below 1000 bp. Nick translation, the conventionally used enzymatic DNA labeling method in CGH, leads to even shorter fragments often below a critical limit for successful analysis. In this study we report the alternative application of non‐enzymatic, PHOTOPROBE® biotin labeling for conjugation of the hapten to the DNA prior to in situ hybridization and fluorescence detection. We analyzed 51 FFPE tumor samples mainly from the upper respiratory tract by both labeling methods. In 19 cases, both approaches were successful. The comparison of hybridized metaphases showed a distinct higher fluorescence signal of the PHOTOPROBE® samples sometimes with a discrete cytoplasm background which however did not interfere with specificity and sensitivity of the detected chromosomal imbalances. For further 32 cases characterized by an average DNA fragment size below 1000 bp, PHOTOPROBE® biotin was the only successful labeling technique thus offering a new option for CGH analysis of highly degraded DNA from archival material. |
| Databáze: | OpenAIRE |
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