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About this book
Experts present methods and protocols essential for understanding parasites at the molecular level. The protocols cover culturing techniques for the major experimental organisms, methods for isolating and processing nucleic acids and proteins, PCR-based protocols for parasite identification, gene isolation and mutation, antibody-based procedures, chromosome and epitope mapping, flow cytometry, RNA sequencing, and parasite transformation.
Contents
Culturing and Biological Cloning of T. brucei. Culturing and Biological Cloning of T. cruzi. In Vitro Cultivation and Biological Cloning of Leishmania. Simple In Vitro Cultivation of the Malaria Parasite P. falciparum (Erythrocytic Stages) Suitable for Large-Scale Preparations. Synchronization and Cloning of Malaria Parasites. The Culture and Prparation of Gametocytes of Plasmodium falciparum for Immunochemical Molecular and Mosquito Infectivity Studies. The Culture of S. mansoni and Production of Life Cycle Stages. Preparation of DNA and RNA from T. brucei. Isolation of RNA and DNA from T. cruzi. Isolation of DNA and RNA from Leishmania. The Extraction and Purification of DNA and RNA from In Vitro Cultures of the Malaria Parasite P. falciparum. Extraction and Purificaiton of Nucleic Acids from Schistosomes. Application of DNA and Whole Organisms to Filter Supports for DNA Probe Analysis: Dot Slot and Touch Blotting. The Development and Use of Repetitive Sequences as DNA Probes for Parasite Detection and Species Identification. Analysis of the DNA of Parasitic Protozoa by Flow Cytometry. PCR for Low-Level Detection of Malaria Parasites in Blood. PCR Methods for Identification of Point Mutations and Gene Rearrangements. Antigenic Typing of Field Isolates of Plasmodium by DNA Techniques. Ribosomal RNA Probes for Detection and Identification of Species. Rapid Sequencing of Parasite rRNA. Antibody Screening of Expression Libraries. Antibody Select Procedure for Characterization of Expression Clones. Isolation of Parasite Genes Using Synthetic Oligonucleotides. Chromosome Mapping Methods for Parasitic Protozoa. Transfection of Leishmania and Trypanosoma brucei by Electroporation. Two-Dimensional Polyacrylamide Gel Electrophoresis. Isoenzyme Electrophoresis for Parasiste Characterization. The Surface Labeling of Schistosomes. Monoclonal Antibody Affinity Chromatography. Immunoblotting and Enzyme Linked Immunosorbent Assay. Immunofluorescence of Parasites. Immunoelectron Microscopic Localization of Antigens in Malaria Parasites. Preparation of Blotted Membrane for Protein Microsequencing. Identification of T- Cell Epitopes. Index.
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