Genetic analyzer (8-Capillary sequencer)

• Subject Area: Medicine
• Term: Funded in 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 538074129

Today, the search for genetic sequence variants is mostly carried out with Next Generation Sequencing (NGS) methods, but direct DNA sequence analysis of individual sequence segments by means of sanger sequencing is still used in research and routine diagnostics. The raw data is read out using special software, and the fine analysis is performed by experienced staff on a computer monitor. DNA sequence analysis is used in routine for mutation search and detection of known mutations in mono- and oligogenic diseases. Capillary sequencing is the gold standard for mutation detection and validation of variants identified by NGS techniques, for example. The most important advantages of direct DNA sequence analysis over screening methods include the comparability of data based on a largely standardized method, the robustness and reproducibility of the method, the assurance of quality through international interlaboratory comparisons, and the comparative ease of performance without the need for extensive optimization steps. Multi-channel (8/16/48/96) capillary electrophoresis instruments are used in routine diagnostics. For many years, the KIM-A research laboratory at UMG has been carrying out investigations of genetic risk factors in gastrointestinal diseases, especially those of the pancreas. These diseases usually have a complex genetic risk, and the contribution of specific genes, or individual sequence variants, is being investigated in current research projects. Therefore, an 8-capillary sequencer is needed for genetic sequence analysis of patient DNA, primarily to investigate potential new risk genes and to identify previously unknown genetic risk variants. For newly identified sequence variants, functional analyses are performed to elucidate the pathomechanism. In addition to the identification of new variants, diagnostics for previously described risk variants are also part of the accurate characterization of patient cohorts, e.g. for further genome-wide association studies. Occurrence and frequency of sequence variants are investigated in patient cohorts and control groups to assess their genetic risk for acute and chronic diseases or severe disease progression, or to assess the complex association of different risk genes, or the genetic risk contribution of potential new risk genes. In further research projects, the sequencer is needed for the generation and control of plasmids and vectors, as well as for genetic studies of transgenic and knock-out constructs or for genotyping of e.g. transgenic or knock-out mouse lines.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Kapillarsequenziergerät (8 Kapillare) für die Sangersequenzierung
• Instrumentation Group: 3150 DNA-Sequenzer
• Applicant Institution: Universität Greifswald
• Leader: Dr. Frank Ulrich Weiss