Comparison of ethylene carbonate and formamide as components of the hybridization mixture in FISH



Título del documento: Comparison of ethylene carbonate and formamide as components of the hybridization mixture in FISH
Revista: Scientia Agricola
Base de datos: PERIÓDICA
Número de sistema: 000455526
ISSN: 0103-9016
Autors: 1
1
1
Institucions: 1University of Szczecin, Faculty of Biology, Szczecin, West Pomerania. Polonia
Any:
Volum: 78
Número: 4
País: Brasil
Idioma: Inglés
Tipo de documento: Artículo
Enfoque: Experimental, analítico
Resumen en inglés The protocols for in situ hybridization (ISH) techniques can vary considerably; however, they usually include denaturation and hybridization steps. Denaturing compounds are used to reduce denaturation and hybridization temperature, which keeps the proper morphology of the preparation. Formamide is the most commonly used reagent in in situ hybridization to lower the melting temperature. The substitution of toxic formamide for a non-toxic ethylene carbonate at 20 % and 50 % concentration in the hybridization mixture helped obtain a high quality in situ hybridization result with two sequences characteristic for rye, JNK, and Bilby. The results after hybridization, with a duration of 90 min and 16 h, were identical when formamide or ethylene carbonate were used in the mixture. In addition, the toxic formamide was eliminated from the post-hybridization steps and specific hybridization signals for both probes were still obtained
Disciplines Agrociencias
Paraules clau: Gramíneas,
Hibridización,
Secale vavilovii,
Carbonato de etileno,
Formamida,
Hibridación fluorescente in situ (FISH)
Keyword: Gramineae,
Hybridization,
Secale vavilovii,
Fluorescence in situ hybridization (FISH),
Ethylene carbonate,
Formamide
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