CONTIG MAPS OF HUMAN CHROMOSOME 17Q22Q23: YAC

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The specific goal is to complete construction of the YAC contig map for about 15 cM sex-average region within the 17q22-q23 (at D17S86 locus: MPO-GH1 region). The specific aims are: (a) define the order of 12 microsatellite markers and size of all (about 25) YACs representing 17q22-q23 region by inter Alu-PCR fingerprinting and Pulsed Field Gel Electrophoresis (PFGE), (b) complete the gaps between different markers by (I) employing one of the techniques e.g. "vectorette"-PCR; to amplify small end-specific fragments of the human DNA insert from the appropriate YACs, (ii) cycle sequence the verified fragment by an enzymatic method using commercially available kits and an automated DNA sequencer, (iii) design PCR primers that are unique for the fragment (Sequence-Tagged-Site; STS) using software e.g. PRIMER or DNASIS, and test the primers on human DNA template, (iv) employ the STSs to obtain overlapping YAC clone(s) from the Michigan Genome Center or CEPH (Centre d'Etude du Polymorphisme Humain), France or through commercial service such as Research Genetics. Other available internal probes such as pC63, pTHH59, pTh17.12, 128E1 and pRMU1 will also be utilized to define more STSs in 17q22-q23 region.