Dark quenchers for donor-acceptor energy transfer

摘要

The present invention provides a family of dark quenchers, termed Black Hole Quenchers (“BHQs”), that are efficient quenchers of excited state energy but which are themselves substantially non-fluorescent. Also provided are methods of using the BHQs, probes incorporating the BHQs and methods of using the probes.

主权项

1. A quencher of excited state energy having the structure:wherein
X and Y are members independently selected from reactive functional groups;
wherein at least one of X and Y is a substituted alkyl amine comprising a reactive functional group selected from carboxy, N-hydroxysuccinimide ester, N-hydroxybenztriazole ester, acid halide, acyl imidazole, thioester, p-nitrophenyl ester, alkenyl ester, alkynyl ester, aromatic ester, —NH2, and phosphoramidite; or
at least one of X and Y is a substituted alkyl amine comprising a covalent bond to a solid support or to a nucleic acid; m is a number selected from 0 to 4, inclusive, such that when m is greater than 1, the X groups are the same or different; v is a number from 1 to 10 and when v is greater than 1, the R2 groups are the same or different; and R1, R2, and R3 are members independently selected from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl and substituted or unsubstituted unsaturated alkyl, with the proviso that at least two of R1, R2 and R3 are members selected from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl and combinations thereof.