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Figure 1 | Chromogenic detection of a biotin-labeled probe.
The Thermo Scientific Biotin Chromogenic Detection Kit is a convenient tool for the chromogenic detection of biotinylated nucleic acid probes. The kit is optimized to reproducibly provide high sensitivity with a low background in applications, such as Southern, Northern, dot and slot blotting, and screening of viral plaques and bacterial colonies. Biotinylation of DNA/RNA probes is widely used as a safe and convenient alternative to radioactive labeling. Biotin can be incorporated into nucleic acids using various enzymatic or non-enzymatic methods, including the Biotin DecaLabel DNA Labeling Kit.
Biotinylated probes are detected with streptavidin coupled to alkaline phosphatase (AP). Streptavidin-AP conjugates bind specifically and irreversibly to the biotin-labeled probes. The probes are then visualized using a chromogenic substrate for alkaline phosphatase BCIP/NBT, which produces a blue-purple precipitate. Visualization does not require X-ray film or other specific equipment (see Figure 1).
The positively charged SensiBlot Plus Nylon Membrane is a recommended component of this detection system to ensure the highest sensitivity of detection and the lowest background.
Figure 2 | Dot-blot hybridization with a biotin-labeled probe.Lambda DNA/HindIII was biotin-labeled with the Biotin DecaLabel DNA Labeling Kit and used as a hybridization probe in a dot-blot of the homologous DNA on SensiBlot Plus Nylon Membrane. The blot was developed with the Biotin Chromogenic Detection Kit.