What We Use
Protein purification is routinely performed on an AKTA FPLC system from GE Healthcare by use of immobilised metal affinity chromatography followed by gel permeation chromatography.
Spectrometry in the laboratory utilises two state-of-the-art instruments from BMG Labtech. We utilise a PHERAStar FS for high-throughput and high-sensitivity UV/Vis and fluorescence spectrometry. We also employ a FLUOStar Omega with an Atmospheric Control Unit to provide a controlled atmospheric environment for bacterial pathogen studies.
For particle analyses we use a Zetasizer Nano ZSP from Malvern Instruments. This instrument allows for the measurement of size, electrophoretic mobility of proteins, zeta potential of nanoparticles, and the microrheology of protein. In our group we utilise it to assess protein quality prior to biochemical and structural characterisation.
Quantitative Real-Time PCR
Analysing gene expression is an essential aspect of a molecular microbiology laboratory and is handled by instrumentation from Thermofisher. The Quantstudio 7 allows us to analyse samples in 96 or 384 well plates quickly and with high levels of repoducibility. The flexible optical platform also allows us to monitor protein unfolding by fluoresence.
High resolution analysis of single gene changes in complex systems is the strength of the Quantstudio 3D Digital PCR system from Thermofisher. This state-of-the-art system allows us to rapidly detect and analyse the frequency of rare gene transcription events in bacterial or eukaryotic samples.
Automated Liquid Handling
The automation of routine tasks, such as DNA minipreps and DNA or RNA isolation, in our laboratory has been a valuable way to improve researcher efficiency, without loss of results quality, and enables them to focus their time on more complex experimental procedures. The QIAcube from Qiagen is an automated liquid handling platform that enables our researchers to have certain routine laboratory procedures performed to a high level of efficiency without the loss of their time to undertake the task.
The incubation and growth of bacterial cells for protein expression is a crucial requirement of a modern laboratory. In our group we utilise the New Brunswick Innova 44R large platform shakers from Eppendorf. These superior instruments allow us to grow large quantities of cells at defined temperatures with high levels of precision and reliabilty.
To extract recombinantly expressed protein from bacterial cells we use a shearing disruption method facilitated by the Constant Systems Cell Disruptor. This instrument breaks cells by firing them at a stainless steel target at up to 800 m/s! This results in highly efficient and rapid cell lysis so that we can obtain our proteins of interest.
We use Beckman Coulter centrifuges. Notably we have recently purchased an Optima MAX-XP ultracentrifuge for small scale preparative experiments (e.g. sucrose-density gradients, small scale membrane protein solubilisation).