Clostridium difficile Methods and Protocols

This second edition provides 21 new chapters on methods used in laboratories for investigating the physiology and molecular genetics of the pathogen Clostridium difficile. Chapters detail up-to -date experimental techniques for gene editing and transcriptional analysis which are used to investigate the fundamental biology of the organism and its virulence factors.  Additional chapters  describe development of potential new treatments including vaccines, bacteriophage and faecal transplantation. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Clostridium difficile: Methods and Protocols, Second Edition provides a comprehensive catalogue of molecular tools and techniques authored by the researchers who have developed them.

Restriction Endonuclease Analysis Typing of Clostridium Difficile Isolates.- Direct PCR-ribotyping of Clostridium difficile.- From FASTQ to Function: In silico Methods for Processing Next Generation Sequencing Data.- Clostridium difficile Genome Editing Using pyrE Alleles.- Use of mCherryOpt Fluorescent Protein in Clostridium difficile.- A Fluorescent Reporter for Single Cell Analysis of Gene Expression in Clostridium difficile.- Clostridium difficile Adhesins.- Intestinal Epithelial Cell Response to Clostridium difficile Flagella.- Isolating and Purifying Clostridium difficile Spores.- Inducing and Quantifying Clostridium difficile Spore Formation.- Characterization of Functional Prophages in Clostridium difficile.- Induction and Purification of C. difficile Phage Tail-like Particles.- Phage Transduction.- Transfer of Clostridium difficile Genetic Elements Conferring Resistance to Macrolide-Lincosamide-streptograminB (MLS_B) Antibiotics.- Methods for Determining Transfer of Mobile Genetic Elements in Clostridium difficile.- Investigating Transfer of Large Chromosomal Regions Containing the Pathogenicity Locus between Clostridium difficile Strains.- An in vitro Model of the Human Colon: Studies of Intestinal Biofilms and lostridium difficile Infection.- MiniBioReactor Arrays (MBRAs) as a Tool for Studying C. difficile Physiology in the Presence of a Complex Community.- A Practical Method for Preparation of Fecal Microbiota Transplantation.- Ion Exchange Chromatography to Analyze Components of a Clostridium difficile Vaccine.- A Size Exclusion Chromatography Method for Analysis of Clostridium difficile Vaccine Toxins.