TRAC 22: Hybridization Techniques: Labeling, Detection and Applications
This 4 day course is designed to introduce the participant to molecular hybridization and in situ hybridization techniques. The application of these techniques to current research questions in genetics and gene expression, molecular pathology, and pathogen detection and identification will be discussed. Probe application and detection systems will serve as the basis for both RNA and DNA in situ hybridization techniques to be addressed in lecture and laboratory. This course will be staffed by clinical and basic scientists familiar with the applications of hybridization techniques to the problems of human disease.
Topics: Probe Design, Optimization, and Application at the Chemical and Cytological Levels; Principles of Hybridization; Base Structure, Modification, Stability and Specificity; Hybridization Kinetics; Preparation of Chemical and Cellular Samples; Hybridization Formats: Target Abundance vs. Probe Abundance, Spots/ Blots, Southern and Northern Analysis, Soluble Targets, in situ Hybridization; Radioactive, Biotinylated, DNP, Chemoluminescent and Oligonucleotide Probe Synthesis, Purification and Use; Label and Signal Amplification and Hybridization Accelerators. DNA in situ hybridization (FISH); Interphase Molecular Cytogenetics; RNA in situ Hybridization; RT-PCR in situ Hybridization; Principles of Karyotyping and Metaphase Chromosome Preparation; Tissue and Specimen Preparation Fixatives, Comparative Genome Hybridization; Spectral Karyotyping; Fluorescence Microscopy; and in situ PCR.