Molecular Probes™ ChromaTide™ dye labeled dUTP, OBEA-dCTP, and UTP nucleotides can be used to synthesize labeled DNA probes without the need for hazardous and expensive radioisotope-labeled nucleotides. These nucleotides can be incorporated using standard molecular biology techniques; labeled probes can then be used in in situ hybridization, microarray, or blotting protocols. ChromaTide™ dye labeled nucleotides are available in different fluorescent colors to facilitate multicolor analysis.

ChromaTide™ Labeled Nucleotides Specifications:
• Ex/Em of dye: Alexa Fluor™ 488-5-UTP (490/520 nm)
• Length of alkynylamino linker: 5 atoms

Methods for Incorporating ChromaTide™ Nucleotides Into Probes
• Nick translation
• Random primer labeling
• End-labeling with terminal deoxynucleotidyl transferase
• Reverse transcription
• PCR amplification

See Methods for Enzymatic Incorporation of ChromaTide™ dUTPs for specific guidelines for each of these methods.

Alexa Fluor™ and BODIPY™ Fluorescent Dyes Make Excellent Probes
Probes made with labeled nucleotides can be used for multicolor techniques such as in situ hybridization and hybridization to arrays. Our proprietary BODIPY™ and Alexa Fluor™ dye conjugates are exceptionally bright, photostable, and essentially pH insensitive. The narrow emission profile of the BODIPY™ dyes helps ensure minimal spectral overlap. The Alexa Fluor™ dyes are highly water soluble, as are DNA probes made from them, making them the labels of choice for fluorescence in situ hybridization.

Long Linkers Improve Performance
The ChromaTide™ dUTP and UTP nucleotides are modified at the C-5 position of uridine via a unique alkynylamino linker, which provides a spacer between the nucleotide and the dye to reduce interactions between them. The number in the product name, e.g., the “12” in fluorescein-12-dUTP, indicates the net length of the spacer, in atoms. These spacers result in brighter conjugates and increased hapten accessibility for secondary detection reagents.

For complete listing of our ChromaTide™ Reagents: Molecular Probes ChromaTide™ and aha labeled nucleotides—Table 8.5.
For additional information on these labeling reagents, read Labeling Oligonucleotides and Nucleic Acids—Section 8.2 in the Molecular Probes™ Handbook.

For Research Use Only. Not intended for human or animal therapeutic or diagnostic use.