Simple and fast in situ hybridization

• Type: Article in International Scientific Journal
• Date: 2002

Authors

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

Journal

Title: Plant Molecular Biology Reporter

Vol. 20

Pages: 219-229

ISSN: 0735-9640

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 5 Citations

Scopus - 7 Citations

Scientific classification

FOS: Natural sciences > Biological sciences

Other information

Authenticus ID: 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 11