Cooperative primers, probes, and applications thereof
Disclosed are compositions and a method relating to amplifying and detecting nucleic acids.
1. A method for synthesizing a nucleic acid, the method comprising:
a. contacting a target nucleic acid with
a cooperative nucleic acid molecule comprising:
i. a first nucleic acid sequence, wherein the first nucleic acid sequence is complementary to a first region of a target nucleic acid, and wherein the first nucleic acid sequence is extendable on the 3′ end;
ii. a second nucleic acid sequence, wherein the second nucleic acid sequence is complementary to a second region of the target nucleic acid, such that it hybridizes to the target nucleic acid downstream from the 3′ end of the first nucleic acid sequence;
iii. a linker connecting said first and second nucleic acid sequences in a manner that allows both the said first nucleic acid sequence to hybridize to the target and extend on the 3′ end and the second nucleic acid sequences to hybridize to the target at the same time;
b. and providing conditions appropriate for nucleic acid synthesis wherein the polymerase extends from the 3′ end of the first nucleic acid sequence through the second nucleic acid sequence, thereby synthesizing a nucleic acid.
2. The method of claim 1 wherein the cooperative nucleic acid molecule comprises a label.
3. The method of claim 1 , wherein more than one cooperative nucleic acid molecule with different sequences are provided.
4. The method of claim 1 , wherein the first nucleic acid molecule will not hybridize to the target without the second nucleic acid molecule hybridizing to the target.
5. The method of claim 1 , wherein the second nucleic acid molecule will not hybridize to the target without the first nucleic acid molecule hybridizing to the target.
6. The method of claim 1 , wherein neither the first nor the second nucleic acid molecule will hybridize to the target without the other hybridizing to the target.
7. The method of claim 1 , wherein effective melting temperature (Tm) of the first nucleic acid molecule is increased by at least 1° C. as compared to the Tm of an isolated first nucleic acid sequence without the second nucleic acid sequence attached to it.
8. The method claim 1 , wherein the second nucleic acid sequence comprises a label.