ABSTRACT
Since cDNA microarrays in general consist of thousands of clones, manual PCR is not applicable. We have used an automated PCR setup consisting of a robotic system from Tecan (Genesis Workstation 200, Tecan, Crailsheim, Germany) combined with a temperature-controlled hotel (automatic incubator cytomat 6002, Heraeus, Hanau, Germany). Template microtiter plates containing bacterial clones as well as empty PCR microtiter plates were stored in the hotel at 4°C (Figure 5.2B, C).The process was set up as follows: microtiter plates from the hotel were placed onto the robotic platform (Figure 5.2A) by means of an elevator within the hotel and a plate gripper of the robot. Next, PCR master mix (50 μl/well) was distributed and bacterial template from glycerol stocks (2 μl/well) was added by a liquid handling system (Figure 5.2A). The finished plates were transported back into the hotel (Figure 5.2B, C). They could be stored there or taken out while the process continued. Ready-to-cycle PCR plates were either immediately processed using external PrimusHT multiblock thermal cyclers (MWG Biotech, Munich, Germany; Figure 5.2D) or stored at −20°C until PCR amplification.
