Compositions for detecting target sequences
View Patent ↗The present invention relates to methods, kits and compositions suitable for the improved detection, quantitation and analysis of nucleic acid target sequences using probe-based hybridization assays.
1. A labeled probe comprising;
a) a clamp, wherein the clamp comprises two PNA segments linked by a flexible linker wherein the clamp hybridizes to a detection sequence present in a third nucleobase segment to thereby form a triplex;
b) at least one donor moiety linked to one PNA segment; and
c) at least one acceptor moiety linked to the other PNA segment.
2. The probe of claim 1 , wherein the donor and acceptor moieties are linked to an end of the first and second PNA segments, respectively.
3. The probe of claim 2 , wherein the donor and acceptor moieties are linked at a position internal to the first and second PNA segments, respectively.
4. The probe of claim 2 , wherein the third nucleobase segment comprises PNA.
5. The probe of claim 2 , wherein the third nucleobase segment comprises DNA or RNA.
6. The probe of claim 5 , wherein the third nucleobase segment is one of two strands of double-stranded DNA.
7. The probe of claim 2 , wherein the third nucleobase segment comprises at least two detection sequences.
8. A PNA probe for hybridizing to a detection sequence, said PNA probe consisting essentially of;
a) two PNA segments connected by a linker;
b) at least one donor moiety linked to one PNA segment; and
c) at least one acceptor moiety linked to the other PNA segment;
wherein the PNA segments can hybridize to the detection sequence to form a triplex, such that the donor and acceptor moieties participate in fluorescence resonance energy transfer to thereby indicate the presence of the preselected detection sequence.
9. The PNA probe of claim 8 , wherein the donor and acceptor moieties are linked to an end of the first and second PNA segments, respectively.
10. The PNA probe of claim 8 , wherein the donor and acceptor moieties are linked at a position internal to the first and second PNA segments, respectively.
11. The PNA probe of claim 8 , wherein the preselected detection sequence comprises PNA.
12. The PNA probe of claim 8 , wherein the preselected detection sequence comprises DNA or RNA.
13. The PNA probe of claim 12 , wherein the preselected detection sequence is one of two strands of double-stranded DNA.
14. The PNA probe of claim 8 , wherein the preselected detection sequence comprises at least two detection sequences.