Main methodological focus
The main focus of the laboratory is the epigenetic characterization of DNA samples. For the analysis of methylation patterns in the promoter region of certain genes, bisulfite sequencing (according to Sanger) has been used in the last 10 years. Recently, nanopore sequencing has been established in our laboratory to analyze larger DNA regions using Cas9-mediated enrichment. This method also allows us to sequence the whole genome. Once different methylation patterns are detected in a clinical picture, the functional relevance or the effect on expression level is checked by real-time PCR, Western blot, ELISA and further gene expression analyses and is correlated with psychometric data. Thereby establishing the correlations between the regulation of relevant genes and the expression of psychologically conspicuous behavioral patterns.
The Laboratory for Molecular Neuroscience is used by many researchers of the department. There is a particularly close cooperation to the "Molecular Psychiatry Research Group".
Equipment of the laboratory
The laboratory is equipped with two Biomek Nxp robots from Beckman-Coulter. Due to the versatile programmability of these auxiliary robots, the following processes can be performed:
- Purification of DNA and RNA from sample material and
- Preparation and post-processing of PCRs and sequencing PCRs,
- Routine pipetting of large clinical / experimental samples
In all these processes, automation achieves more consistent and stable results compared to manual processing and a higher throughput.
For DNA analysis we have two sequencing methods available:
Nanopore sequencing has been established in our laboratory for the analysis of base sequence and methylation patterns of larger DNA regions using Cas9-mediated enrichment or the entire genome without prior Bisulphite chemical conversion and PCR amplification.
Further DNA analyses (bisulfite sequencing, SNP, VNTR, LPR) by means of Sanger sequencing are carried out on an ABI 3500xL sequencer from Applied Biosystems.
For the amplification of nucleic acids we have four thermal cyclers and one real-time cycler from BioRad at our disposal. For the visualization of the PCR products we use an electrophoresis system (chambers and PowerPac™ Basic power supply) and GelDoc Imaging System from BioRad.
The DNA-/RNA/protein quantification is carried out by spectrophotometer / fluorometer of the series DS-11 of the company Denovix.
The Bioanalyzer 2100 of the company Agilent is used for the more exact determination of size and quality of DNA and RNA.
For the concentration of low concentrated (DNA-) samples our laboratory is equipped with the Savant DNA 120 Speedvac Concentrator from Thermo Scientific.
Protein detection and protein quantification is performed using protein and blot chambers, VersaDoc imaging system from BioRad and ELISA equipment from Molecular Devices including plate washer from BioTek.
For the cultivation of cell lines, primary cells and iPS cells we have a fully equipped S1 laboratory with sterile workbenches, incubators, centrifuge, microscope and live cell imager.
For cloning and expression of recombinant proteins as well as restriction, ligation and transformation, two rooms of our S1 laboratory are equipped for genetic engineering work at safety level 1. CRISPR/Cas9 technology, gene expression and promoter activity studies using the GloMax Multi Detection System from Promega and genotyping are also used here. For these methods, incubators from Memmert, a temperature-controlled shaker from Edmund Bühler GmbH including incubation hood for the cultivation of (recombinant) microorganisms and an electroporator for transfection are available.
For the storage of sample material, the laboratory has five ultra-low freezers from the company Sanyo. The sample material can be sustainably managed by means of the software MySamples by storing it in drawer inserts of the company Landgraf especially suitable for the low temperature range.
Leaders of the laboratory
Prof. Dr. med. Helge Frieling
Deputy director & Founding member and scientific advisory board of the Research Core Unit Genomics of the MHH
Phone: +49 511 532 7275
Fax: +49 511 532 7276
Prof. Dr. med. Stefan Bleich
Director of the Department of Psychiatry, Social Psychiatry and Psychotherapy
Phone: +49 511 532 6748