Methods of labelling biomolecules with fluorescent dyes

Abstract

Fluorescent energy transfer cassettes that allow through bond energy transfer and have a succinimidyl ester functionality suitable for affecting them to biomolecules or provided and are applied to high throughput DNA sequencing.

We claim:  
     
         1 . A fluorescent dye for labelling a biological molecule comprising: 
 a donor group, said donor group comprising a UV absorbing chromophore;    at least one acceptor group, said acceptor group comprising a fluorescent emitter, wherein said UV chromophore absorbs energy at a lower wavelength than that emitted by the fluorescent emitter, and said donor group and said acceptor group are conjugated to each other such that energy transfer between the donor group and the acceptor group comprises transfer via a through bond mechanism.    
     
     
         2 . The dye of  claim 1  wherein said donor group is an anthracene derivative.  
     
     
         3 . The dye of  claim 1  wherein said acceptor group is a BODIPY fragment.  
     
     
         4 . The dye of  claim 1  further comprising a functionality group for attaching said dye to the biomolecule.  
     
     
         5 . The dye of  claim 4  wherein said functionality group comprises a bromine functionality on the donor group for attachment to the biomolecule and said biomolecule comprises a DNA molecule.  
     
     
         6 . The dye of  claim 4  wherein said functionality group comprises a succinimidyl ester.  
     
     
         7 . The dye of  claim 1  further comprising a functional group for incorporating a label on said dye.  
     
     
         8 . The dye of  claim 1  wherein said chromophore absorbs energy at about 266 nanometers.  
     
     
         9 . The dye of  claim 1  further comprising a plurality of fluorescent emitters, each with a different emission wavelength.  
     
     
         10 . The dye of  claim 1  further comprising a second chromophore wherein said UV absorbing chromophore and said second chromophore absorb in mutually exclusive regions of the spectrum.  
     
     
         11 . The dye of  claim 1  wherein said biological molecules are labelled for high throughput DNA sequencing.  
     
     
         12 . A method for labelling a biological molecule comprising contacting the molecule with the dye of  claim 1.