Patent ID: 6316230
Filing Date: 2001-11-13
Classification: C12Q

Abstract:
A method of producing a non-radioisotopically labelled chimeric extension product comprising the step of enzymatically extending a PNA-DNA chimera in the presence of a template nucleic acid, a polymerase and a primer extension reagent, wherein said primer extension reagent comprises a nucleotide 5'-triphosphate capable of effecting enzymatic chimera primer extension, the PNA-DNA chimera or the nucleotide 5'-triphosphate is labelled with a non-radioisotopic label selected from the group consisting of a fluorescent dye, a fluorescence quencher, a hybridization stabilizer, an energy-transfer dye pair, an electrophoretic mobility modifier, a chemiluminescent dye, an amino acid, a protein, a peptide, an enzyme, and an affinity ligand, andthe PNA-DNA chimera has the structure:P.sub.x --L--N.sub.y 3'wherein:each P is independently a PNA monomer;x is an integer from 5 to 15;L represents a covalent linkage between P and N;each N is independently a nucleotide; andy is an integer from 3 to 15;with the proviso that the 3' terminal N has a 3' hydroxyl group.