This detailed collection covers methods recently developed for experiments on organisms from each of the major euglenozoan groups, including diplonemids, euglenids, and kinetoplastids. Volume 1 presents a historical review of the research into euglenozoans, followed by sections of techniques involving isolation and culturing of different species of euglenozoans, as well as a variety of -omics research methods. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, Euglenozoa: Methods and Protocols, Volume 1 serves as an ideal guide for researchers working with these fascinating organisms and investigating the aspects of their extreme biology.
Chapter 14 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Why Euglenozoans.- Isolation and Cultivation of Diplonemids.- Preparation of Diplonemid Samples for Microscopy.- Protocol for the Molecular Identification of Autotrophic Euglenids in Environmental Samples.- Culture and Genetic Modification of the Monoxenous Trypanosomatids Crithidia fasciculata and Crithidia bombi.- Poly(A) Profiling for Euglenozoans Using Oxford Nanopore Direct RNA Sequencing, Illustrated by the Example of Euglena gracilis.- Euglenid Extrachromosomal DNA: Assembly and Annotation.- Nuclear Genome Assembly and Annotation of Kinetoplastids.- Assembly and Annotation of Kinetoplastid and Diplonemid Mitochondrial Genomes.- Cleavage Under Targets and Release Using Nuclease (CUT&RUN) for Trypanosoma brucei Parasites.- Mapping Open Chromatin in Trypanosoma brucei Using ATAC-Seq.- Transcriptomic Analysis of Trypanosomatids.- Small RNA Profiling in Trypanosomes.- Desorption Electrospray Ionization–Mass Spectrometry Imaging (DESI-MSI) of Metabolites in Heart Samples Infected with Trypanosoma cruzi.- Microbiome Profiling in Chagas Disease: Sample Collection, Sequencing, and Analysis.- Metabolomics Analysis of Trypanosomes Using Ion Mobility-Enhanced Mass Spectrometry.- Exploring Protein Interactomes Using TurboID-Directed Proximity Labeling and Mass Spectrometry.- Mapping Trypanosoma Protein Interactome by Proximity-Dependent Biotinylation.- From Genome to Function: Metabolism-Wide Models of Lifecycle Stages of Parasites.- Enhanced Detection of Homology Using Artificial Intelligence in Euglenids.
