The current PCR protocol in the Olink assay consists of 40 amplification cycles and has a total run time of 2 h 10 min. Since this step, next after the overnight incubation, is the most time-consuming step of the reaction,
Olink started an investigation to find out whether a shortening of this protocol would be possible. An evaluation of the key features for the negative control samples run on each sample plate; blank
buffer samples that represent background values with high Ct, showed that the maximum Ct value measured during validation experiments was 30.4. Based on this information, an investigation on the possibility to reduce the number of PCR-cycles to 35 and thereby save 10 min run time was initiated.