Abstract：
: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.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Clausson CM,Arngården L,Ishaq O,Klaesson A,Kühnemund M,Grannas K,Koos B,Qian X,Ranefall P,Krzywkowski T,Brismar H,Nilsson M,Wählby C,Söderberg Odoi
10.1038/srep12317subject
Has Abstractpub_date
2015-07-23 00:00:00pages
12317issn
2045-2322pii
srep12317journal_volume
5pub_type
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