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Compaction of rolling circle amplification products increases signal integrity and signal-to-noise ratio
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2015 (English)In: Scientific Reports, ISSN 2045-2322, Vol. 5, 12317Article in journal (Refereed) Published
Abstract [en]

Rolling circle amplification (RCA) for generation of distinct fluorescent signals in situ relies upon the self-collapsing properties of single-stranded DNA in commonly used RCA-based methods. By introducing a cross-hybridizing DNA oligonucleotide during rolling circle amplification, we demonstrate that the fluorophore-labeled RCA products (RCPs) become smaller. The reduced size of RCPs increases the local concentration of fluorophores and as a result, the signal intensity increases together with the signal-to-noise ratio. Furthermore, we have found that RCPs sometimes tend to disintegrate and may be recorded as several RCPs, a trait that is prevented with our cross-hybridizing DNA oligonucleotide. These effects generated by compaction of RCPs improve accuracy of visual as well as automated in situ analysis for RCA based methods, such as proximity ligation assays (PLA) and padlock probes.

Place, publisher, year, edition, pages
2015. Vol. 5, 12317
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Other Biological Topics
URN: urn:nbn:se:kth:diva-171886DOI: 10.1038/srep12317ISI: 000358358900001PubMedID: 26202090ScopusID: 2-s2.0-84937944116OAI: diva2:846074
Science for Life Laboratory - a national resource center for high-throughput molecular bioscienceSwedish Research Council

QC 20150814

Available from: 2015-08-14 Created: 2015-08-10 Last updated: 2015-08-14Bibliographically approved

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Brismar, Hjalmar
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Cell PhysicsScience for Life Laboratory, SciLifeLab
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