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® 546-16-OBEA-dCTP (555/570 nm)
• Length of alkynylamino linker: 16 atomsMethods 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.