title: Quantification of polymerase chain reaction products by affinity-based hybrid collection.
abstract: We have used oligonucleotides modified with biotin in the 5'-end as primers in the polymerase chain reaction (PCR)-amplification. This results in the synthesis of 5'-biotinylated DNA molecules, which are detected by hybridization to a labelled probe in solution. The formed hybrids are collected on an avidin-matrix by mediation of the biotin residue of the target molecules. The affinity-based hybrid collection method is quantitative and makes it possible to measure the amount of DNA produced in the PCR-amplification. At low concentrations of template the efficiency of the process is close to 100%, making it possible to detect the presence of a few molecules of target DNA in 25 cycles. With high template concentrations the efficiency of the process is low.
authors.affiliation: Orion Genetic Engineering Laboratory, Orion Corporation Ltd, Helsinki, Finland.
authors.forename: A C
authors.initials: AC
authors.lastname: Syvänen
authors.forename: M
authors.initials: M
authors.lastname: Bengtström
authors.forename: J
authors.initials: J
authors.lastname: Tenhunen
authors.forename: H
authors.initials: H
authors.lastname: Söderlund
mesh_terms: D000345:Affinity Labels; D001483:Base Sequence; D001710:Biotin; D003587:Cytomegalovirus / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase; D005784:Gene Amplification; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D019914:Taq Polymerase
chemical_list: D000345:Affinity Labels; D004279:DNA, Viral; D015345:Oligonucleotide Probes; D001710:Biotin; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase