Digital quantification of rolling circle amplified single DNA molecules in a resistive pulse sensing nanopore
2015 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 67, no SI, 11-17 p.Article in journal (Refereed) Published
Novel portable, sensitive and selective DNA sensor methods for bio-sensing applications are required that can rival conventionally used non-portable and expensive fluorescence-based sensors. In this paper, rolling circle amplification (RCA) products are detected in solution and on magnetic particles using a resistive pulse sensing (RPS) nanopore. Low amounts of DNA molecules are detected by padlock probes which are circularized in a strictly target dependent ligation reaction. The DNA-padlock probe-complex is captured on magnetic particles by sequence specific capture oligonucleotides and amplified by a short RCA. Subsequent RPS analysis is used to identify individual particles with single attached RCA products from blank particles. This proof of concept opens up for a novel non-fluorescent digital DNA quantification method that can have many applications in bio-sensing and diagnostic approaches.
Place, publisher, year, edition, pages
2015. Vol. 67, no SI, 11-17 p.
Padlock probe, RCA, Single molecule detection, Resistive pulse sensing, Nanopore
IdentifiersURN: urn:nbn:se:uu:diva-248795DOI: 10.1016/j.bios.2014.06.040ISI: 000350076900003PubMedID: 25000851OAI: oai:DiVA.org:uu-248795DiVA: diva2:801885