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. 2017 Apr 6;8:14738. doi: 10.1038/ncomms14738

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Copyright © 2017, The Author(s)

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(a) ‘Multi-colour' imaging of phiX174 with multiple shape IDs and their distance analysis. Left, three different shape IDs, triangular (corresponding to site 1,433), triangular with STVs (corresponding to site 1,529) and cross (corresponding to site 4,914), are site-specifically labelled on phiX 174 with different labelling efficiency (85% for the single-site labelling, 56.4% for the two-site labelling and 30.8% for the three-site labelling). Scale bar, 200 nm. Right, histograms for the counts of shape IDs as a function of distance between site 1,433, 1,529 and 4,914 in phiX. As for two-site labelling (left), the measured distance between site 1,433 and 4,914 is 650 nm, which is in good agreement with calculated distance 648 nm. As for three-site labelling (right), the measured distances between site 1,433, 1,529 and 4,914 are 652 nm (blue) and 682 nm (red), which are in good agreement with the calculated distances 648 and 680 nm. Also see Supplementary Figs 14 and 15. (b) Schematic showing and three-dimensional AFM topographic images of triangular- (corresponding to site 1,433), cross- (corresponding to site 1,463) and STV-decorated triangular- (corresponding to site 1,529) shaped IDs for labelling phiX 174. Left, the contour length between two labelled sites (1,433 and 1,463) is ∼10 nm (30 bp). Right, the contour length between two labelled sites (1,433 and 1,529) is ∼32 nm (96 bp).