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1990•Nucleic Acids ResearchOpen access

Non-radioactive labeling of RNA transcriptsin vitrowith the hapten digoxigenin (DIG); hybridization and ELISA-based detection

Hans-Joachim Höltke, Christoph Keßler

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Abstract

We have developed a system for the enzymatic in vitro synthesis of non-radioactively labeled RNA which is derivatized with the hapten digoxigenin (DIG). The labeling reaction as well as the conditions for hybridization and detection of hybrids by an antibody-conjugate and a coupled colour reaction were analyzed and adapted for high sensitivity and low background. In addition, data on the performance and sensitivity of digoxigenin-labeled RNA probes in Southern and Northern blots are presented.

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What this paper is about

We have developed a system for the enzymatic in vitro synthesis of non-radioactively labeled RNA which is derivatized with the hapten digoxigenin (DIG). The labeling reaction as well as the conditions for hybridization and detection of hybrids by an antibody-conjugate and a coupled colour reaction were analyzed and adapted for high sensitivity and low background. In addition, data on the performance and sensitivity of digoxigenin-labeled RNA probes in Southern and Northern blots are presented.

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Available abstract

We have developed a system for the enzymatic in vitro synthesis of non-radioactively labeled RNA which is derivatized with the hapten digoxigenin (DIG). The labeling reaction as well as the conditions for hybridization and detection of hybrids by an antibody-conjugate and a coupled colour reaction were analyzed and adapted for high sensitivity and low background. In addition, data on the performance and sensitivity of digoxigenin-labeled RNA probes in Southern and Northern blots are presented.

Key concepts: Digoxigenin, Dig, Biology, Hapten, RNA, Molecular biology, Blot, In vitro

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Non-radioactive labeling of RNA transcriptsin vitrowith the hapten digoxigenin (DIG); hybridization and ELISA-based detection — Research Paper | ScholarLens