Non-radioactive labeling of RNA transcripts in vitro with the hapten digoxigenin (DIG); hybridization and ELISA-based detection.
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.
Höltke
H J
HJ
Boehringer Mannheim GmbH, Biochemical Research Center, Nonnenwald, Penzberg, FRG.
Kessler
C
C
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D004076:Digoxigenin / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens; D006801:Humans; D007700:Kinetics; D015394:Molecular Structure; D009693:Nucleic Acid Hybridization; D015347:RNA Probes / Q000737:chemistry* / Q000378:metabolism; D014158:Transcription, Genetic
D006241:Haptens; D015347:RNA Probes; D004076:Digoxigenin