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Highly specific DNA detection employing ligation on suspension bead array readout
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Molecular tools. Uppsala University, Science for Life Laboratory, SciLifeLab.
2015 (English)In: New Biotechnology, ISSN 1871-6784, E-ISSN 1876-4347, Vol. 32, no 5, 504-510 p.Article in journal (Refereed) Published
Abstract [en]

We show for the first time that monomerized rolling circle amplification (RCA) products can be directly detected with the Luminex suspension bead array readout without the need of PCR amplification. Furthermore, using monomerized RCA products to guide ligation of the detection oligonucleotide (DO) to barcode sequences on the magnetic Luminex beads, combined with efficient washing and increased measurement temperature, yields a higher signal to noise ratio. As a proof-of-principle, we demonstrate detection of pathogenic DNA sequences with high reproducibility, sensitivity and a dynamic range over four orders of magnitude. Using padlock probes in combination with bead suspension arrays opens up the possibility for highly multiplexed DNA targeting and readout.

Place, publisher, year, edition, pages
2015. Vol. 32, no 5, 504-510 p.
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:uu:diva-256977DOI: 10.1016/j.nbt.2015.01.011ISI: 000355666700010PubMedID: 25681158OAI: oai:DiVA.org:uu-256977DiVA: diva2:838727
Available from: 2015-07-01 Created: 2015-06-29 Last updated: 2017-12-04Bibliographically approved

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Kuhnemund, Malte

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