Duell T, Wang M, Wu J, Kim U J, Weier H U
Life Sciences Division, University of California, Berkeley, California, 94720, USA.
Genomics. 1997 Nov 1;45(3):479-86. doi: 10.1006/geno.1997.4954.
Quantitative DNA fiber mapping (QDFM) allows rapid construction of near-kilobase-resolution physical maps by hybridizing specific probes to individual stretched DNA molecules. We evaluated the utility of QDFM for the large-scale physical mapping of a rather unstable, repeat-rich 850-kb region encompassing the immunoglobulin lambda variant (IGLV) gene segments. We mapped a minimal tiling path composed of 32 cosmid clones to three partially overlapping yeast artificial chromosome (YAC) clones and determined the physical size of each clone, the extent of overlap between clones, and contig orientation, as well as the sizes of gaps between adjacent contigs. Regions of germline DNA for which we had no YAC coverage were characterized by cosmid to cosmid hybridizations. Compared to other methods commonly used for physical map assembly, QDFM is a rapid, versatile technique delivering unambiguous data necessary for map closure and preparation of sequence-ready minimal tiling paths.
定量DNA纤维图谱分析（QDFM）通过将特定探针与单个拉伸的DNA分子杂交，能够快速构建近千碱基分辨率的物理图谱。我们评估了QDFM在对一个相当不稳定、富含重复序列的850 kb区域进行大规模物理图谱绘制中的实用性，该区域包含免疫球蛋白λ可变区（IGLV）基因片段。我们将由32个黏粒克隆组成的最小拼接路径定位到三个部分重叠的酵母人工染色体（YAC）克隆上，并确定了每个克隆的物理大小、克隆之间的重叠程度、重叠群方向，以及相邻重叠群之间间隙的大小。对于我们没有YAC覆盖的种系DNA区域，通过黏粒与黏粒杂交进行了表征。与常用于物理图谱组装的其他方法相比，QDFM是一种快速、通用的技术，可提供图谱封闭和制备序列就绪的最小拼接路径所需的明确数据。
