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Biotin Chromogenic Detection Kit

Store at -20°C

Biotin Chromogenic Detection Kit is a convenient tool for the chromogenic detection of biotinylated nucleic acid probes.
  
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Chromogenic detection of a biotin-labeled probe

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. 

Features

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).

Highlights

  • 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

Applications

  • Southern, Northern, dot/slot blots, plaque, or colony screening.

Includes

  • Streptavidin-AP Conjugate
  • BCIP/NBT Solution
  • Blocking Reagent
  • 10X Blocking/Washing Buffer
  • 10X Detection Buffer
  • Detailed Protocol

Notes

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.

  
Storage Condition-20 C
HazardousNo
Quality ControlThe kit is functionally tested in dot-blot hybridization with biotin-labeled probe. 0.03 pg of homologous DNA is detected.
Chromogenic detection of a biotin-labeled probe

Chromogenic detection of a biotin-labeled probe


Dot-blot hybridization with a biotin-labeled probe

Dot-blot hybridization with a biotin-labeled probe

Dot-blot hybridization with a biotin-labeled probe

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.