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Characterization of four human YAC libraries for clone size, chimerism and X chromosome sequence representation.
Authors:R Nagaraja  J Kere  S MacMillan  M J Masisi  D Johnson  B J Molini  G R Halley  K Wein  M Trusgnich  and B Eble
Institution:Center for Genetics in Medicine, Washington University School of Medicine, St Louis, MO 63110.
Abstract:Four collections of human X-specific YACs, derived from human cells containing supernumerary X chromosomes or from somatic cell hybrids containing only X human DNA were characterized. In each collection, 80-85% of YAC strains contained a single X YAC. Five thousand YACs from the various libraries were sized, and cocloning was assessed in subsets by the fraction of YAC insert-ends with non-X sequences. Cocloning was substantial, ranging up to 50% for different collections; and in agreement with previous indications, in all libraries the larger the YACs, the higher the level of cocloning. In libraries made from human-hamster hybrid cells, expected numbers of clones were recovered by STS-based screening; but unexpectedly, the two collections from cells with 4 or 5 X chromosomes yielded numbers of YACs corresponding to an apparent content of only about two X equivalents. Thus it is possible that the DNA of inactive X chromosomes is poorly cloned into YACs, speculatively perhaps because of its specialized chromatin structure.
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