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RAPID NONRADIOACTIVE IN-SITU HYBRIDIZATION FOR INTERLEUKIN-2 MESSENGER-RNA WITH RIBOPROBES GENERATED USING THE POLYMERASE CHAIN-REACTION JOURNAL OF IMMUNOLOGICAL METHODS Birk, P. E., Grimm, P. C. 1994; 167 (1-2): 83-89

Abstract

In situ hybridization is a technique with widespread application. However, its usefulness has been limited by the need for radioactive materials and the requirement for the DNA to be cloned onto an appropriate vector. We have utilized the polymerase chain reaction to directly incorporate a T7 RNA polymerase promoter sequence onto the cDNA for interleukin-2. Digoxigenin-labelled riboprobes were then synthesized using this PCR product as a template. The digoxigenin-labelled riboprobes were then used in non-radioactive in situ hybridization to detect messenger RNA for interleukin-2 in mitogen stimulated peripheral blood mononuclear cells. This methodology has the potential for widespread application in immunology and cytokine research.

View details for Web of Science ID A1994MU31400009

View details for PubMedID 8308289