CONTIG MAPS OF HUMAN CHROMOSOME 17Q22Q23: YAC

具体目标是完成YAC重叠群的构建 17q22-q23内约15厘米性别平均区的地图(在D17S86 基因座：MPO-GH1区)。具体目标是：(A)界定秩序 12个微卫星标记和所有(约25个)YAC的大小 用Alu-PCR间隔区指纹图谱表示17q22-q23区域 脉冲场凝胶电泳(PFGE)，(B)填补了 不同的标记通过(I)使用一种技术，例如 “载体”--聚合酶链式反应；扩增人类末端特异的小片段 从适当的YAC插入DNA，(Ii)验证的循环序列 使用市场上可获得的试剂盒的酶方法和 自动DNA测序仪，(Iii)设计独一无二的聚合酶链式反应引物 对于片段(序列标记位点；STS)，使用软件，例如 引物或DNASIS，并在人DNA模板上检测，(Iv) 利用STSS从密歇根获得重叠的YAC克隆(S) 基因组中心或CEPH(人类多态中心)，法国 或通过研究遗传学等商业服务。其他 提供内部探头，如pC63、pTHH59、pTh17.12、128E1和 还将利用pRMU1在17q22-q23区域定义更多的STS。

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.