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Quantification of polymerase chain reaction products by affinity-based hybrid collection
1988 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 16, no 23, p. 11327-11338Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
1988. Vol. 16, no 23, p. 11327-11338
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Medical and Health Sciences
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URN: urn:nbn:se:uu:diva-169909PubMedID: 2849762OAI: oai:DiVA.org:uu-169909DiVA: diva2:508021
Available from: 2012-03-07 Created: 2012-03-07 Last updated: 2017-12-07Bibliographically approved

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Syvänen, Ann-Christine

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