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. 2014 Nov 24;9(11):e112656. doi: 10.1371/journal.pone.0112656

Figure 3. Artifactual amplification is abolished by using an invasion oligonucleotide (IO) with a 2′-O-methyl RNA modification.

Figure 3

(A) Configuration of the IO molecules used. (B) Real-time monitoring of SIBA reactions with SYBR Green I using different IOs: (i) IO with a 2′-O-methyl RNA modification and fully homologous to the target duplex, SB-IO; (ii) IO fully homologous to the target duplex, where the 2′-O-methyl RNA modification was replaced with natural DNA nucleotides, SB-IO DNA; (iii) IO with a 2′-O-methyl RNA modification that is not homologous to the target duplex, SB-IO DIFF-METH; and (iv) IO with the 2′-O-methyl RNA modification deleted, SB-IO NON-METH. SB-F21 and SB-R21 were the forward and reverse primers, respectively. The reactions were either performed using 106 target template molecules (SB-template) or in the absence of template.