Simple and fast in situ hybridization

• Tipo: Artigo em Revista Científica Internacional
• Data: 2002

Autores

Borlido, J (Autor) Outra; Pereira, S (Autor) FCUP; Ferreira, R (Autor) Outra; Coelho, N (Autor) Outra; Duarte, P (Autor) Outra; Pissarra, J (Autor) FCUP

Revista

Título: Plant Molecular Biology Reporter

Vol. 20

Páginas: 219-229

ISSN: 0735-9640

Editora: Springer Nature

Indexação

ISI Web of Knowledge - 5 Citações

Scopus - 7 Citações

Classificação Científica

FOS: Ciências exactas e naturais > Ciências biológicas

Outras Informações

ID Authenticus: P-000-N7S

DOI: 10.1007/bf02782457

Abstract (EN): RNA in situ hybridization is a powerful technique but can be time consuming and potentially hazardous. We developed a procedure in which nonisotopic in situ hybridization is performed in Vibrato me-sectioned tissues that are not dehydrated or embedded in paraffin or resin, thus avoiding additional pretreatment steps to allow the probes to penetrate the cell structure. Predigestion with proteases was not necessary. Hybridization of sections and immunologic detection were performed in microplates to avoid the use of slide-coating agents. Biotinylated cDNA probes were synthesized by PCR. Synthetic oligonucleotide probes chemically labeled with biotin or digoxigenin were also used. Stained sections were mounted on slides for microscopic observation, and various transcripts were successfully detected using different visualization methods. This method is fast and can be easily implemented in other laboratories because it requires minimal expertise in processing biological samples for microscopy.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 11