Multi-acceptor molecular probes and applications thereof
AssigneeUniversity of Florida Research Foundation, Inc.
InventorsWeihong Tan, Chaoyong Yang
Life of this patent
Estimating the term…
Abstract
An oligonucleotide-based molecular probe includes at least one pin loop, the pin loop including a loop sequence complementary to a target sequence. A first stem sequence is attached to one end of the pin loop, the first stem having at least one fluorescent label attached thereto. A second stem sequence is attached to the other end of the pin loop. The second stem has a plurality of quencher molecules attached thereto.
Claims21
A molecular probe, comprising:
- at least one pin loop, said pin loop including a loop sequence complementary to a target sequence;
- a first stem sequence attached to one end of said pin loop, said first stem having at least one fluorescent label attached thereto, and
- a second stem sequence attached to the other end of said pin loop, said second stem having a plurality of quencher molecules attached thereto.
A molecular probe comprising:
- an aptamer having both a 5′-end and a 3′-end;
- at least one fluorescent molecule attached to one of the 5′-end and the 3′-end;
- and
- a plurality of quencher molecules attached to the other of the 5′-end and the 3′-end.
A detection system comprising:
- at least one excitation light source providing excitation radiation;
- and
- at least one oligonucleotide-based molecular probe comprising at least one pin loop, said pin loop comprising a loop sequence complementary to a target sequence, a first stem sequence attached to one end of said pin loop, said first stem having at least one fluorescent label attached thereto, and a second stem sequence attached to the other end of said pin loop, said second stem having a plurality of quencher molecules attached thereto.
A method for forming molecular beacons each having a plurality of quenchers, the method comprising the steps of:
- providing at least one fluorophore labeled nucleotide monomer, a plurality of quencher labeled nucleotide monomers, and a plurality of bases;
- and
- coupling a programmed sequence of said at least one fluorophore labeled nucleotide monomer, said plurality of bases, and said plurality of quencher labeled nucleotide monomers.
A method for forming molecular beacons each having a plurality of quenchers, the method comprising the steps of:
- coupling a programmed sequence of bases to form a sequence;
- coupling dendrimeric phosphoramidite to said sequence, wherein said dendrimeric phosphoramidite generates multiple reactive OH groups;
- and
- coupling a plurality of quencher molecules to said sequence at said reactive OH groups.
A method for forming molecular beacons each having a plurality of quenchers, the method comprising the steps of:
- coupling a plurality of quencher molecules to at least one base to form a multiple quencher moiety;
- and
- attaching said multiple quencher moiety to an oligonucleotide comprising at least one pin loop having a first end and a second end, said pin loop including a loop sequence complementary to a target sequence, a first stem sequence attached to the first end of said pin loop, said first stem having at least one fluorescent label attached thereto, and a second stem sequence attached to the second end of said pin loop, said second stem having a plurality of quencher molecules attached thereto.
A method of detecting the presence of at least one target molecule in a sample, the method comprising the steps of:
- providing an aptamer comprising a binding region that binds to the target molecule and both a 5′-end and a 3′-end, a fluorophore attached to one of the 5′-end and the 3′-end, and at least one quencher attached to the other of the 5′-end and the 3′-end;
- providing the sample;
- contacting the aptamer with the sample and allowing binding of the aptamer to said at least one target molecule,
- wherein binding of the at least one target molecule to the aptamer results in either an increase or decrease of fluorescence intensity compared to a baseline fluorescence of unbound aptamer;
- and
- measuring the resultant fluorescence intensity.
18-21. (canceled)
Dependencies are inferred from the claim text.