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. 2019 Jan 1;9(1):279–289. doi: 10.7150/thno.28474

Figure 3.

Figure 3

The evaluation of “unlocked mechanism” and signal enrichment amplification strategy. (A) The principle of the double key unlocking detection assay with and without target miRNAs. The “unlocked mechanism” based on the DSN-miRNA double key and the mechanism of DSN-assisted target recycling and signal amplification are illustrated in the presence of target miRNAs. (B) The effect of signal enrichment events triggered by the “unlocked mechanism”. FCM detection and fluorescent images of PS-ssDNA before (no fluorescence) and after (with fluorescence) reactions. (C) The “unlocked mechanism” only works well on the target miR-21 and DSN, forming the DSN-miRNA double key. The concentration of miR-21 was 1 nM. (D) Agarose gel electrophoresis (3%) used for the exploration of the DSN cleaving function on DNA/RNA dimers. The left lane is the DNA ladder marker (20 bp to 200 bp). Lane #1 is HDs+ miRNA, lane #2 is HDs + miRNA + DSN, lane #3 is HDs, lane #4 is miRNA and lane #5 is HDs + DSN. (E) Fluorescent confocal images of the Cy5 signal in the homogeneous phase and anchored on the PS. (F) The fluorescent intensity of the Cy5 signal in the homogeneous phase and on the microbeads by a signal enrichment strategy. Error bars show standard deviations (n = 3).