On-chip solid-phase rolling-circle amplification (spRCA) system for testing proximity ligation assay (PLA) templates.

<p>(A) The image shows a merged fluorescence and bright-field micrograph of two circular pull-down areas within representative unit cells on the chip. In the lower and upper images, positive and negative RCA events were obtained with matched and mismatched shared connectors (SCs), respectively. The bar graph shows background corrected fluorescence signal corresponding to the pull-down area of the left images. Error bars are calculated from four repeats. The scale bar represents 75 µm. (B) Temperature dependence of a PLA template assembling in a spRCA reaction. The graphs show the local background corrected fluorescence signals of 160 spRCA reactions, including the screening of one CC/DC pair, of 100 binary adaptor combinations from the selected 10 PLA templates. Furthermore, 20 single adaptors, 20 non-specific DNA strands, and 20 tests without DNA were screened against the same connector pair as control.