Digital quantification of rolling circle amplified single DNA molecules in a resistive pulse sensing nanopore.

K├╝hnemund M, Nilsson M

Biosens Bioelectron 67 (-) 11-17 [2015-05-15; online 2014-06-24]

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.

Affiliated researcher

PubMed 25000851

DOI 10.1016/j.bios.2014.06.040

Crossref 10.1016/j.bios.2014.06.040

pii: S0956-5663(14)00453-9

Publications 9.5.0