Fret process

A pair of FRET hybridization probes hybridizing adjacently to a target nucleic acid sequence, comprising

• a first member of said pair of FRET hybridization probes comprising:
• a nucleotide sequence entity which is substantially complementary to the sequence of the target nucleic acid;
• a fluorescent entity, said entity being a FRET donor entity;
• a first spacer entity connecting said nucleotide sequence entity and said first fluorescent entity;
• a second member of said pair of FRET hybridization probes comprising
• a nucleotide sequence entity which is substantially complementary to the sequence of the target nucleic acid;
• a second fluorescent entity, said second fluorescent entity being a FRET acceptor entity;
• a second spacer entity connecting said nucleotide sequence entity and said fluorescent entity, said second spacer entity being different from said first spacer entity;
• wherein the length of said first spacer entity and the length of said second spacer entity differ in size at least by a connecting chain of 15 atoms.

A pair of FRET hybridization probes hybridizing adjacently to a target nucleic acid sequence comprising:

• a the first member of said pair of FRET hybridization probes comprising:
• a nucleotide sequence entity which is substantially complementary to the sequence of the target nucleic acid;
• a fluorescent entity, said entity being a FRET donor entity;
• a spacer entity connecting said nucleotide sequence entity and said fluorescent entity, said spacer entity comprising a number of n1=1-15 nucleotide residues non-complementary to the target DNA;
• the second member of said pair of FRET hybridization probes comprising:
• a nucleotide sequence entity which is substantially complementary to the sequence of the target nucleic acid;
• a fluorescent entity, said entity being a FRET acceptor entity;
• a spacer entity connecting said nucleotide sequence entity and said fluorescent entity, said spacer entity comprising a number of n2=1-15 nucleotide residues non-complementary to the target DNA;
• wherein the value of n1 differs from the value of n2 by a natural number between 1 and 10.

A set of at least three oligonucleotides, comprising:

• a first oligonucleotide and a second oligonucleotide, said first oligonucleotide and said second oligonucleotide being capable of acting as a pair of amplification primers for a template dependent nucleic acid amplification reaction, each of said first oligonucleotide and said third oligonucleotide being labeled with one corresponding member of a FRET pair consisting of a FRET donor entity and a FRET acceptor entity;
• one of said first said oligonucleotide or said third oligonucleotide comprising:
• a nucleotide sequence entity which is substantially complementary to the sequence of the target nucleic acid;
• a fluorescent entity, said entity being either the FRET donor entity or the FRET acceptor entity;
• a spacer entity connecting said nucleotide sequence entity and said fluorescent entity;
• wherein said spacer entity comprises a connecting chain of at least 15 atoms.

A method for chemical solid phase synthesis of multiple oligonucleotides comprising:

• a) preparation of a dye labeled CPG-(N)n, wherein
• N is arbitrarily chosen nucleotide residues different from G, and n=1-10;
• b) performing a first solid phase synthesis of a first oligonucleotide having a first sequence using the CPG prepared in step a);
• and
• c) performing a second solid phase synthesis of at least a second oligonucleotide having a second sequence using the CPG prepared in step a).