CONTIG MAPS OF HUMAN CHROMOSOME 17Q22Q23: YAC

具体目标是完成YAC重叠群的构建 在17 q22-q23（D17 S86）内约15 cM性别平均区域的图谱 基因座：MPO-GH 1区）。 具体目标是：（a）确定秩序 12个微卫星标记和所有（约25个）YAC的大小 通过Inter-PCR指纹图谱代表17 q22-q23区域， 脉冲场凝胶电泳（PFGE），（B）完成 通过（I）采用以下技术之一， “vectorette”-PCR;扩增人免疫球蛋白的小末端特异性片段， （ii）来自适当YAC的DNA插入物， 使用市售试剂盒通过酶促方法进行片段化， 自动DNA测序仪，（iii）设计独特的PCR引物 对于片段（序列标记位点; STS），使用软件， 引物或DNASIS，并在人DNA模板上测试引物，（iv） 使用STS从Michigan获得重叠的YAC克隆 基因组中心或CEPH（Centre d 'Etude du Polymorphisme Humain），法国 或通过商业服务，如研究遗传学。 其他 可用的内部探针如pC63、pTHH 59、pTh17.12、128 E1和 pRMU 1也将用于在17 q22-q23区域中定义更多的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.