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Isolation and characterization of DNA probes from the short arm of the human X chromosome which detect restriction fragment length polymorphisms Starr, Terence

Abstract

The isolation and characterization of DNA probes which detect RFLPs from a small region of the human genome, Xp, has been the focus of this thesis. Thirty X specific DNA probes were isolated from a flow sorted library constructed from a 49,XXXXY cell line. These probes were confirmed to be from the X chromosome by hybridization to dot blots containing DNA from mouse, human, and a somatic cell hybrid which contained the X chromosome as its only human DNA. Hybridization to somatic cell hybrids containing different regions of the X chromosome localized 9 of the probes to Xp. After testing 185 probe-enzyme combinations three of the Xp probes (localized to the distal 2/3 of Xp) were found to detect RFLPs. One probe, pTS316, was shown not to be inherited in a Mendelian fashion. The presence of one of the polymorphic bands detected by this probe correlated with the phenomenon of X chromosome inactivation.

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