Oligo-riboprobes |
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Authors: | P. Denny Q. Hamid J. E. Krause J. M. Polak S. Legon |
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Affiliation: | (1) Department of Histochemistry and Chemical Pathology, Royal Postgraduate Medical School, Hammersmith Hospital, Du Cane Road, W12 0HS London, UK;(2) Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, USA |
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Abstract: | Summary In situ hybridisation detection of mRNAs using riboprobes has become a widely used technique. However, the identification of cells producing closely-related yet distinct mRNAs is difficult with the usual size probes. More-over, it is not always easy to obtain the required cDNA essential for cRNA probe synthesis. To avoid these problems, we have used synthetic oligodeoxynucleotides to generate short, single stranded RNA probes (oligo-riboprobes). These probes can be labelled to very high (109 cpm/g) specific activity and can be prepared for any published nucleotide sequence. We have used these probes to localise (preprotachykinin) PPT mRNA producing neurons in rat hypothalamus and bowel. The results were compared to that obtained with cRNA probes generated from preprotachykinin cDNA. |
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