Buxton J, Davies J, Shelbourne P, Yokobata K, Williamson R, Johnson K
Department of Anatomy, Charing Cross and Westminster Medical School, London, UK.
Mol Cell Probes. 1993 Feb;7(1):75-80. doi: 10.1006/mcpr.1993.1010.
We describe a method for rapidly isolating overlapping bacteriophage clones corresponding to the genomic region cloned in a yeast artificial chromosome (YAC) that does not require sub-cloning or lambda DNA preparation. Purified YACs from 19q13.3 were used to screen a flow-sorted chromosome 19 library, and the resulting positive clones were characterized using inter-Alu PCR. In addition, aliquots of the lambda stocks were gridded out, and hybridized with probes known to be present in the YACs, thereby avoiding having to perform DNA preparations. The application of this technique in the identification of lambda clones which span the myotonic dystrophy (DM) locus on 19q13.3 is presented, and its general advantages are discussed.
我们描述了一种快速分离与克隆于酵母人工染色体（YAC）中的基因组区域相对应的重叠噬菌体克隆的方法，该方法不需要亚克隆或λDNA制备。从19q13.3纯化的YAC用于筛选经流式细胞仪分选的19号染色体文库，并用Alu间PCR对所得阳性克隆进行表征。此外，将λ噬菌体原液等分试样铺板，并与已知存在于YAC中的探针杂交，从而避免了进行DNA制备。本文介绍了该技术在鉴定跨越19q13.3上强直性肌营养不良（DM）基因座的λ克隆中的应用，并讨论了其一般优势。
