Hsu Joanne H, Zeng Hui, Lemke Kalistyn H, Polyzos Aris A, Weier Jingly F, Wang Mei, Lawin-O'Brien Anna R, Weier Heinz-Ulrich G, O'Brien Benjamin
Department of Molecular and Cell Biology, University of California, Berkeley (UCB), Berkeley, CA 94720, USA.
Int J Mol Sci. 2012 Dec 20;14(1):57-71. doi: 10.3390/ijms14010057.
Chromosome enumeration in interphase and metaphase cells using fluorescence in situ hybridization (FISH) is an established procedure for the rapid and accurate cytogenetic analysis of cell nuclei and polar bodies, the unambiguous gender determination, as well as the definition of tumor-specific signatures. Present bottlenecks in the procedure are a limited number of commercial, non-isotopically labeled probes that can be combined in multiplex FISH assays and the relatively high price and effort to develop additional probes. We describe a streamlined approach for rapid probe definition, synthesis and validation, which is based on the analysis of publicly available DNA sequence information, also known as "database mining". Examples of probe preparation for the human gonosomes and chromosome 16 as a selected autosome outline the probe selection strategy, define a timeline for expedited probe production and compare this novel selection strategy to more conventional probe cloning protocols.
使用荧光原位杂交（FISH）对间期和中期细胞中的染色体进行计数，是一种用于细胞核和极体的快速准确细胞遗传学分析、明确性别鉴定以及定义肿瘤特异性特征的既定方法。该方法目前的瓶颈在于，可用于多重FISH分析的商业化非同位素标记探针数量有限，且开发其他探针的成本相对较高、难度较大。我们描述了一种基于公开可用DNA序列信息分析（即“数据库挖掘”）的快速探针定义、合成及验证的简化方法。以人类性染色体和作为选定常染色体的16号染色体的探针制备为例，概述了探针选择策略，确定了加快探针生产的时间表，并将这种新型选择策略与更传统的探针克隆方案进行了比较。
Zotero 插件
