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).
• High sensitivity of detection—0.03 pg of homologous DNA in dot blot hybridization with biotin-labeled probe • Low background due to optimized washing and blocking procedures • Non-radioactive detection visualization does not require X-ray film or other specific equipment (see Figure 2) • Fast and convenient—ready-to-use components
• Southern, Northern, dot/slot blots, plaque, or colony screening.
Note:This kit is optimized for hybridization procedures using nylon membranes. The detection sensitivity may vary depending on the membranes supplied by different vendors.