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Nucleic Acids Research, 2001, Vol. 29, No. 8 e42
© 2001 Oxford University Press

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

Catherine D. van Raamsdonk and Shirley M. Tilghman1,*

Howard Hughes Medical Institute and 1Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA

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.

* To whom correspondence should be addressed. Tel: +1 609 258 2900; Fax: +1 609 258 3345; Email: stilghman{at}molbio.princeton.edu


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