Optimizing the detection of nascent transcripts by RNA fluorescence in situ hybridization | Zendy

Catherine D. Van Raamsdonk | Zendy

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Optimizing the detection of nascent transcripts by RNA fluorescence in situ hybridization

Author(s) -

Catherine D. Van Raamsdonk

Publication year - 2001

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/29.8.e42

Subject(s) - biology , fluorescence in situ hybridization , in situ hybridization , rna , gene , gene expression , microbiology and biotechnology , genetics , transcription (linguistics) , intron , dna , linguistics , philosophy , chromosome

An unusual feature of the mammalian genome is the number of genes exhibiting monoallelic expression. Recently random monoallelic expression of autosomal genes has been reported for olfactory and Ly-49 NK receptor genes, as well as for Il-2, Il-4 and Pax5. RNA fluorescence in situ hybridization (FISH) has been exploited to monitor allelic expression by visualizing the number of sites of transcription in individual nuclei. However, the sensitivity of this technique is difficult to determine for a given gene. We show that by combining DNA and RNA FISH it is possible to control for the hybridization efficiency and the accessibility and visibility of fluorescent probes within the nucleus.

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