A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storage.
A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storage.
复制标题
3.5 基因组等效多存取 YAC 文库：构建、表征、筛选和存储。
作者：
R. Anand;J. Riley;R. Butler;J. Smith;A. Markham
从人淋巴母细胞样（48）细胞系DNA中构建了35，000个克隆的酵母人工染色体（YAC）初级网格文库。平均YAC大小约为350 kb，代表基因组覆盖率大于3.5倍。将文库在-70 ℃下储存为浸渍有20%甘油的尼龙过滤器上的网格化克隆，并在微量滴定板中储存为单个克隆的甘油悬浮液，从而提供延长的多用户潜力。到目前为止，我们已经使用了14个单拷贝探针，通过菌落杂交以及PCR筛选该文库，并为每个探针分离了1至5个YAC克隆。
The construction of a yeast artificial chromosome (YAC) primary gridded library of 35,000 clones from human lymphoblastoid (48,XXXX) cell line DNA is described. The average YAC size is approximately 350kb representing a greater than 3.5 times coverage of the genome. The library is stored at -70 degrees C as gridded clones on nylon filters impregnated with 20% glycerol and as glycerol suspensions of individual clones in microtitre plates providing a prolonged multi-user potential. To date we have used 14 single copy probes to screen this library by colony hybridisation as well as PCR and have isolated between 1 and 5 YAC clones for every probe.