Morf, N V; Sulzer, A; Kratzer, A; Bär, W (2011). Internal validation of Tecan robots (Freedom EVO® 150 and 75) for PCR and capillary electrophoresis setup. Forensic Science International: Genetics Supplement Series, 3(1):e89-e90.
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We validated and established an automated method for PCR and capillary electrophoresis setup (CE setup), using the Tecan Freedom EVO® 150 and 75 automated liquid handling workstations powered by Freedom EVOware®. The PCR setup script of the Freedom EVO® 150 was based on the HID EVOlution™ System. However, substantial changes in the HID script were required before putting the system on stream in the routine. The CE setup of the Freedom EVO® 75 was based on a script made by application specialists of Tecan. For internal validation of the two robots, we investigated repeatability, reproducibility and occurrence of contaminations. The analysis of the reproducibility data demonstrated that electropherograms of samples pipetted by the robots had a lower mean size standard peak height, than the hand-pipetted samples, whereas the mean sample peak height did not differ between the two methods. This was mainly due to precision-differences between the robots and the hand-pipetting. The investigation of the occurrence of contaminations showed that with the PCR setup robot no sample got contaminated. The CE setup robot on the contrary revealed carryovers of samples from one well to the subsequent well. After intense examination, we discovered that the tip travel height was set too low, causing the tip to touch the rim of the 96 well plate which led to the carryovers. The contact between rim and tip could not be seen by naked eye, but after increasing the tip travel height, no further contamination was observed. Our validation demonstrates that robots, even when set up by application specialists, need to undergo a proper internal validation before being put on stream in the routine.
|Item Type:||Journal Article, not refereed, original work|
|Communities & Collections:||04 Faculty of Medicine > Institute of Legal Medicine|
610 Medicine & health
|Deposited On:||11 Mar 2012 08:50|
|Last Modified:||23 Nov 2012 13:09|
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