In the frame of the ITN Clospore, we propose a skills training for PhD students on Bacterial RNAseq experiments.
Using total RNAs, we will i) enrich the mRNAs by depleting the rRNAs; ii) prepare and sequence cDNA library and iii) analyse and manage sequencing data.
The bacterial endospore, one of the most highly resistant life-forms on earth allows the bacterium to survive exposure to extremes of temperature, desiccation, radiation, disinfectants and, in the case of anaerobic clostridia, oxygen. Whilst the pathogenesis of its notorious pathogens (C. botulinum, C. perfringens, C. difficile) is generally ascribed to the devastating toxins produced (neuro-, endo- and cytotoxins), their capacity to produce spores lies at the heart of the diseases they cause. Spores play a pivotal role in the spread of infection (C. difficile) and in food poisoning (C. botulinum and C. perfringens). The processes of spore formation and germination (return to vegetative cells) represent key intervention points. The majority of clostridia are benign and can sustainably produce useful chemicals and fuels. Crucially, the regulation of chemical production and sporulation is intimately linked. Despite the tremendous importance of spores, little is known of the developmental processes of clostridial sporulation and germination. Deriving this knowledge including the regulatory networks and thence exploiting it, is the objective of CLOSPORE