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. 1988 Dec 9;16(23):11327-38.
doi: 10.1093/nar/16.23.11327.

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

A C Syvänen  1 , M Bengtström, J Tenhunen, H Söderlund
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Free PMC article

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

A C Syvänen et al. Nucleic Acids Res. .
Free PMC article

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.

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