The molecular biology laboratory (Biolab) is one of the core facilities in use from the structural biology group in Trieste. The preparation of the protein samples used in our structural and functional investigations, are carried out at the Biolab. The facility allows for the amplification, manipulation and the mutagenesis of DNA, which is then introduced in specific bacterial cell lines derived from E. coli. Bacteria are grown, the recombinant proteins are expressed and then isolated and purified by using chromatographic methods. The optimization of the storage conditions is the last step of the preparation of the highly pure protein samples, suitable for structural analysis. At the Biolab, several vectors are available for prokaryotic expression as well as several different cell lines, which can be explored in order to gain an optimized expression level of the recombinant protein under study. It is also possible to express recombinant Seleno-Metionine labelled proteins, for their use in crystallographic phasing(SAD/MAD). Many different chromatographic columns (affinity, ion exchange, gel-permeation, idrophobic interactions) are available, for optimized purification protocols. A cold-room at 4° C is also available for unstable proteins especially sensitive to higher temperature.
FPLC Akta PURE
FPLC Akta Prime plus
PCR Eppendorf Mastercycler
Spettrophotometer Thermo-Fisher Nanodrop
Spettrophotometer Pharmacia Ultrospec 3000
Incubator/shaker N-Biotek NB-205 VL (large volumes)
Incubator/shaker Edmund Buhler KS-15 control (small volumes)
Centrifuge Beckman-Coulter J6-MC
Sterile Hood (classe 2)
Other available facilities: low-temperature freezers (-80° e -40° C), refrigerators (4° C), pH-meter, thermoblock, top-table centrifuges (variable temperature also), trans-illuminator, complete apparatus for gel-electrophoresis (both nucleic acids and proteins).
Name and Surname: SONIA COVACEUSZACH
Sede: S.S. 14, Km 163,5 Basovizza – Trieste
Tel: +39 040 3757526
Mail : sonia.covaceuszachATts.ic.cnr.it