Jan Christoph Müller
Titel der Masterarbeit:
Systematic Analyses of Workflow Associated PCR Contamination in a Molecular Laboratory
The laboratory of the Department of Historical Anthropology of the Georg-August-University of Goettingen successfully works with Polymerase-Chain-Reaction (PCR)-based DNA analysis for more than 20 years. Two of the main research projects of the department are the investigations of ancient and forensic specimens by the use of Short Tandem Repeats (STR) analysis via PCR. Since the very beginning of DNA analysis in the department, thousands of PCR reactions have been performed. Thus, a huge variety of PCR products (amplicons) must have been generated over the past decades. In the light of contamination prevention the question raises, what traces of these longtime work processes can be tracked down today.
It is known, that amplicons can easily disperse by aerosol droplets within a laboratory. These droplets can form trough air movement, for example by the opening of reaction tubes, after a PCR. Small droplets, with a volume of just 0.005µL, can contain millions of copies as potential template DNA and therefore represent a huge contamination risk for DNA and especially for ancientDNA laboratories. Hence, the aim of this thesis is the analysis of amplicon-aerosol dispersion within the facility, as well as the investigation of aerosol cloud forming by the use of fluorescent dye.