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Nucleic Acids Research, 1988, Vol. 16, No. 23 11327-11338
© 1988

CHEMISTRY

Quantification of polymerase chain reaction products by affinity-based hybrid collection

Ann-Christine Syvänen, Marina Bengtström, Tenhunen Jukka and Hans Söderlund

Orion Genetic Engineering Laboratory, Orion Corporation Ltd Valimotie 7, SF-00380 Helsinki, Finland

Received September 30, 1988. Revised November 2, 1988. Accepted November 2, 1988.

We have used oligonucleotides nodified 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.


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