Patent ID: 11926870
Assignee: AGILENT TECHNOLOGIES, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 4:
5. A method of performing Polymerase Chain Reaction (PCR) comprising amplifying a template comprising a target sequence with the following reagents:
1) a Polymerase Chain Reaction (PCR) primer comprising the following general structure:, wherein:
5′ Acceptor: represents one member of a fluorescence resonance energy transfer (FRET) pair;
A: represents a nucleic acid sequence motif of 10 to 30 bases;
3-C: represents a linker region comprised of at least three carbon atoms;
B: represents a double stranded restriction enzyme recognition site;
A′: represents a nucleic acid sequence motif of 10 to 30 bases being complementary to the nucleic acid sequence motif of A; and
R-prim: represents a nucleic acid sequence complementary to a target sequence in a nucleic acid sequence to be amplified;
2) a PCR primer S-prim providing, in combination with the PCR primer as defined in 2), amplification of at least one amplicon comprised in said target sequence;
3) a donor oligonucleotide having the following general structure:, wherein
A′ represents a nucleic acid sequence motif of 10 to 30 bases being complementary to the nucleic acid sequence motif of A;

4) a restriction enzyme; and
5) amplification enzyme and nucleotides; and
6) template comprising a target sequence and amplifying the template using the PCR primer.