| This thesis describes development and application of two independent approaches of fluorescence in situ hybridization using cyanine dye-labeled DNA probes.;In the second project, synthesis and purification of Cy3-4- and Cy5-4-dUTP analogs are described. It is demonstrated that Cy3-4-dUTP and Cy5-4-dUTP are efficient substrates for DNA polymerases. They can be incorporated into DNA probes by nick translation, random priming and polymerase chain reaction. Important parameters are optimized to give labeling densities in the range of 20-40 dyes per kilobase of DNA. Multicolor FISH on chromosomes and interphase nuclei with cyanine dye-labeled DNA probes are demonstrated.;In the first project, chemical labeling of oligonucleotide probes with cyanine dyes is described. A rapid and sensitive procedure of fluorescence in situ hybridization (FISH) to fixed cells in suspension is developed using these 5... |
