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. 2014 Apr 22;9(4):e95192. doi: 10.1371/journal.pone.0095192

Figure 1. Design and description of 96-plex PEA.

Figure 1

(A) 94 pairs of specific antibodies are equipped with oligonucletotides (PEA probes) and mixed with an antigen-containinig sample. (B) Upon sample incubation, all proximity probe pairs bind their specific antigens, which brings the probe oligonucleotides in close proximity to hybridize. The oligonucleotides have unique annealing sites that allows pair-wise binding of matching probes. Addition of a DNA polymerase leads to an extension and joining of the two oligonucleotides and formation of a PCR template. (C) Universal primers are utilized to preamplify all 96 different DNA templates in paralell. (D) Uracil-DNA glycosylase partly digests the DNA templates and remove all unbound primers. (E) Finally each individual DNA sequence is detected and quantified using specific primers in by microfluidic qPCR.