Highly specific DNA detection employing ligation on suspension bead array readout
2015 (English)In: New Biotechnology, ISSN 1871-6784, Vol. 32, no 5, 504-510 p.Article in journal (Refereed) Published
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.
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
IdentifiersURN: urn:nbn:se:uu:diva-256977DOI: 10.1016/j.nbt.2015.01.011ISI: 000355666700010PubMedID: 25681158OAI: oai:DiVA.org:uu-256977DiVA: diva2:838727