Workflow for detection of ligands using nucleic acids

A method for detecting a target in a sample wherein:

• a) binding a first and a second probe, each of which binds specifically to the target, wherein each of the probes comprises an oligonucleotide portion or tail;
• b) ligating the first and second oligonucleotide tails thereby producing a ligated oligonucleotide template;
• and,
• c) performing a polymerase chain reaction (PCR) of the oligonucleotide template across the first and second oligonucleotide to quantify the said template.

A method for detecting a target in sample comprising:

• a) binding a first and a second probe, wherein each probe binds specifically to the target, each of said probes comprise an oligonucleotide portion or tail;
• b) ligating the oligonucleotides to produce a ligated oligonucleotide template and amplifying the template by PCR in a single step to produce an amplified template;
• and,
• c) quantitating the amplified template

The method of claim 24 or 25, wherein said ligase is selected from the group consisting of a nucleic acid ligase, oligonucleotide ligase, DNA ligase, T3 DNA ligase, T4 DNA ligase, T5 DNA ligase, T7 DNA ligase, vaccinia virus DNA ligase, E. coli DNA ligase, mammalian DNA ligase I, mammalian DNA ligase II, mammalian DNA ligase III, Tth DNA ligase, KOD DNA ligase, a thermostable DNA ligase, RNA ligase, T4 RNA ligase, bacteriophage RB69 RNA ligase, Autographa californica nuclearpolyhedrosis virus RNA ligase, a thermophilic RNA ligase, bacteriophage RM378 RNA ligase, bacteriophage TS2126 RNA ligase, a small-footprint DNA ligase (SFL), the ligase of SEQ ID NO.:

• 77, the ligase of SEQ ID NO.:
• 78, the ligase of SEQ ID NO.:
• 79, the ligase of SEQ ID NO.:
• 80, the ligase of SEQ ID NO.:
• 81, the ligase of SEQ ID NO.:
• 82, a derivative thereof, a fragment thereof, and combinations thereof.