Mapping of Low-Frequency Chimeric Yeast Artificial Chromosome Libraries from Human Chromosomes 16 and 21 by Fluorescence in Situ Hybridization and Quantitative Image Analysis

摘要: Abstract Yeast artificial chromosome (YAC) clones from low-frequency chimeric libraries of human chromosomes 16 and 21 were mapped onto diploid fibroblast metaphase using fluorescence in situ hybridization (FISH) digital imaging microscopy-YACs selected to provide subregional location ordering known unknown markers on the long arm 21, particularly Down syndrome region (q22). YACs overlap regions spanning cosmid maps. YAC indirectly labeled with fluorescein, total DNA was counterstained propidium iodide. A single image containing both FISH signal whole acquired for each interest fluorescent probe a spread. From digitized image, intensity profile through axis gave length position. The map position expressed as fractional (FL) relative end short (FLpter). clone hybridized, 20-40 images analyzed. Thirty-eight 16, their FLs distributed along arms. On 47 mapped, including 12 markers. To confirm order dense population within region, two-color mapping strategy used which an anonymous located one or two chromosome. Our FL maps have 1- 2-Mb resolution, measurement has typical standard error 0.5-1 Mb.