Key responsibilities
:
• To assist in research aimed at understanding nucleo-cytoskeletal coupling.
• To express and purify proteins from bacteria and insect cell lines.
• To characterise the purified proteins and protein complexes using biochemical and biophysical methods.
• To culture and transfect human cell lines.
• To perform immunostaining and immunoblotting experiments.
• To perform fluorescent microscopy of human cell lines using widefield and confocal instruments.
• To assist in preparing samples for cryo-EM and cryo-ET
• Keep detailed and accurate records.
• To ensure the research is carried out in accordance with good practice and in compliance with local policies and legal requirements.
• To plan and manage your experimental work according to the objectives set by your supervisor.
• To actively develop and troubleshoot challenging assays.
Working relationships:
You will report to Dr Piotr Kolata and Dr Allegretti. You will interact with other members of the group, and with other scientists at the LMB as necessary.
Additional information:
The above lists are not exhaustive, and you are required to undertake such duties as may reasonably be requested within the scope of the post. All employees are required to act professionally, co-operatively and flexibly in line with the requirements of the post and the MRC.
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Education / qualifications / training required:
Essential: Bachelor’s degree in Biochemistry, Cell or Molecular Biology or a related subject.
Desirable: Research Master’s degree in a related biological subject.
Previous work experience required/ technical skills:
The candidate must have several of the essential skills listed below.
Essential:
• DNA construct design and cloning, including Gibson assembly, restriction cloning, bacteria transformation, and plasmid DNA isolation.
• Expression and purification of recombinant proteins from bacteria and/or insect cell lines using affinity, ion exchange, and/or size-exclusion chromatography.
• Experience in using automated protein purification systems.
• Biochemical characterisation of proteins: SDS-PAGE, western blotting
• Mammalian cell culture, including passaging and transfections.
• Immunofluorescence.
• Light microscopy using widefield and confocal systems.
Desirable:
• Working with challenging protein complexes.
• Biophysical characterisation of protein complexes (Isothermal Titration Calorimetry, Surface Plasmon Resonance, fluorescent activity assays, mass photometry, negative-stain transmission electron microscopy)
• Knowledge of bioinformatics tools SnapGene, BLAST, ChimeraX, PyMol, ImageJ, AlphaFold
• Independence in design and optimisation of experiments relevant to the post, including troubleshooting.
Knowledge and experience:
Essential:
• The ability to work safely without close supervision.
• The ability to follow complex protocols.
• Good communication skills to allow accurate information to be exchanged.
• Excellence in keeping lab records.
Desirable:
• Able to acquire new techniques quickly and independently.
Personal skills / behaviours / qualities:
Essential:
• Highly organised and detail-oriented individual.
• Willingness to independently develop, troubleshoot, and validate new methods or pipelines.
• A mature and conscientious attitude.
• Skills and experience in working collaboratively as a member of a team.
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