Quantification of polymerase chain reaction products by affinity-based hybrid collection
- 1 January 1988
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 16 (23) , 11327-11338
- https://doi.org/10.1093/nar/16.23.11327
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.This publication has 21 references indexed in Scilit:
- IDENTIFICATION OF HIV-INFECTED SERONEGATIVE INDIVIDUALS BY A DIRECT DIAGNOSTIC TEST BASED ON HYBRIDISATION TO AMPLIFIED VIRAL DNAThe Lancet, 1988
- Transcription of the dystrophin gene in human muscle and non-muscle tissuesNature, 1988
- Direct Detection of HIV-1 RNA from AIDS and ARC Patient SamplesDNA, 1988
- Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA PolymeraseScience, 1988
- Detection of human papilloma virus in paraffin-embedded tissue using the polymerase chain reaction.The Journal of Experimental Medicine, 1988
- [21] Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reactionPublished by Elsevier ,1987
- Fast quantification of nucleic acid hybrids by affinity-based hybrid collectionNucleic Acids Research, 1986
- Non-radioactive hybridization probes prepared by the chemical labelling of DNA and RNA with a novel reagent, photobiotinNucleic Acids Research, 1985
- Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase IJournal of Molecular Biology, 1977
- Repeated Sequences in DNAScience, 1968