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).
- 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.
- Streptavidin-AP Conjugate
- BCIP/NBT Solution
- Blocking Reagent
- 10X Blocking/Washing Buffer
- 10X Detection Buffer
- Detailed Protocol
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.
|Quality Control||The kit is functionally tested in dot-blot hybridization with biotin-labeled probe. 0.03 pg of homologous DNA is detected.|
|Storage Condition||-20 C|
|Storage Condition:||-20 C|
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