The Biochemistry Core Facility is the institute’s central unit devoted to scientific services. We are organized under the office of the Managing Director and our methods portfolio covers the following areas:
DNA sequencing from plasmids and PCR products or ready-to-load samples is performed. We use a modified Sanger dideoxy terminator cycle sequencing chemistry, the ABI BigDye kit version 3.1, on a ABI 3730 48-capillary sequencer and 36 cm capillaries.
Automated solid phase synthesis of peptides of 4 to ca. 60 amino acids using Fmoc/tBu-chemistry. Characterization and quality control is done by RP-HPLC (purity) and MS (identity) and most common modifications are available. In addition, an amino acid analysis (relative amino acid composition) is available.
The Chemistry Service offers the synthesis, purification and characterization of organic compounds (substrates, inhibitors, fluorophores, conjugates, caged compounds etc.).We can also advise you concerning your chemical problems, and will help you in chemical research in databases investigations for the development of new synthetic routes for desired target molecules.
A set of biophysical methodologies for studying protein structure and folding, protein-protein or protein-ligand interactions and protein complex formation is provided as open-access instruments or full service. CD spectroscopy, microcalorimetry, dynamic light-scattering, analytical ultracentrifugation, surface plasmon resonance (Biacore) and ESI-TOF mass spectrometry are available, among others.
Recombinant Protein Production
The service for protein expression and purification is assisting projects in designing strategies for high level expression of soluble proteins and by providing vectors and strains from our in-house collection. Specifically, we clone the gene of interest into expression vectors, rapidly screen for optimal combinations of vectors, strains and conditions and produce the protein in E. coli and insect cells up to 10 l scale. A multiple fermentation system is also available, as well as fermentation of organisms like S. cerevisiae or P. pastoris. Proteins are then purified using standard chromatography techniques and quality control is assessing protein identity, purity, stability and folding.
Custom OligoRNA Synthesis Service
The custom oligoRNA synthesis service provides chemically synthesized oligoribonucleotides in the size range between 10 to about 45 nucleotides which could be used in RNA interference (RNAi) experiments for example.
Mass Spectrometry Service
The Mass Spectrometry Core Facility will routinely provide services including protein identification of Coomassie & silver-stained gel bands and solution samples; post-translational modification mapping of Coomassie-stained gel bands of purified proteins (phosphorylation, ubiquitinylation, acetylation, etc.), and relative peptide/protein quantification by stable isotope labeling and label free methods (SILAC) using highly sensitive nanoLC/MS/MS techniques by a fast scanning and highly sensitive 2D linear ion trap and a high resolution/high mass accuracy LTQ-Orbitrap mass spectrometer.