Lengauer C, Henn T, Onyango P, Francis F, Lehrach H, Weith A
Research Institute of Molecular Pathology, Vienna, Austria.
Genet Anal Tech Appl. 1994;11(5-6):140-7. doi: 10.1016/1050-3862(94)90034-5.
A series of 80 microclone probes derived from the chromosomal region 1p36 was used to isolate corresponding clones from the ICRF human P1 library (see Francis et al., this issue). Hybridization screenings were performed using probe pools on high-density filter grids. A total of 87 P1 clones specific for 1p36 were isolated. This large-scale approach allowed a detailed evaluation of the complexity, quality, and utility of this library. The isolated P1 clones were used both for size determination by pulsed-field gel electrophoresis and as probes for fluorescence in situ hybridization (FISH) analysis. FISH of P1 clones is shown to be both easy and efficient to perform on metaphase chromosomes and interphase nuclei. This observation is expected to reveal new avenues for diagnosis of disease-related chromosomal changes. The use of P1 clones as a tool in clinical and tumor interphase cytogenetics is discussed and compared with FISH data of other long insert clones such as cosmids and YAC clones.
使用一系列源自染色体区域1p36的80个微克隆探针，从ICRF人类P1文库中分离出相应的克隆（见本期Francis等人的文章）。使用探针池在高密度滤膜网格上进行杂交筛选。共分离出87个对1p36特异的P1克隆。这种大规模方法使得能够对该文库的复杂性、质量和实用性进行详细评估。分离出的P1克隆既用于通过脉冲场凝胶电泳确定大小，也用作荧光原位杂交（FISH）分析的探针。结果表明，P1克隆在中期染色体和间期核上进行FISH操作既简便又高效。这一发现有望为诊断与疾病相关的染色体变化开辟新途径。本文讨论了将P1克隆用作临床和肿瘤间期细胞遗传学工具的情况，并与其他长插入片段克隆（如黏粒和酵母人工染色体克隆）的FISH数据进行了比较。
