In situ hybridisation of EBV DNA-DNA hybrids using wet heat in polypropylene containers
| Autor: | L G Labrecque |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
In situ
Polypropylene Herpesvirus 4 Human Hot Temperature Microwave oven General Medicine In situ hybridization Biology Nucleic Acid Denaturation Polypropylenes Molecular biology Pathology and Forensic Medicine chemistry.chemical_compound chemistry Virology DNA Viral Denaturation (biochemistry) Low copy number DNA In Situ Hybridization Research Article |
| Zdroj: | Journal of clinical pathology. 45(12) |
| ISSN: | 0021-9746 |
| Popis: | AIMS: To explore procedures designed to optimise DNA-DNA in situ hybridisation, using cells infected with Epstein-Barr virus (EBV) and tissues and subfragments of the EBV DNA as probes. METHODS: The denaturation step occurred in a polypropylene container, using wet heat generated by a hot water container, the pressure cooker, or the microwave oven, without coverslips, reaching a temperature of 121 degrees C or more in these two last systems. Two different visualisation systems were used. RESULTS: Fixed cells and tumours harbouring a high and medium to low copy number (a few hundreds to 33 copies per cell), were clearly labelled, using a simple reiterated subfragment (BamW) of the EBV DNA, and fresh frozen cells, harbouring a very low copy number (one to two on average) labelled using BamW as well as BamH (single non-reiterated 6 kilobase subfragment). CONCLUSION: This is a valuable alternative technique for DNA-DNA ISH that can be used in fresh frozen samples as well as fixed samples. |
| Databáze: | OpenAIRE |
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