Manitoba Institute of Cell Biology, Cancer Care Manitoba, University of Manitoba, 675 McDermot Avenue, Winnipeg, MB, R3E 0V9, Canada.
Cytotechnology. 2004 Mar;44(3):143-9. doi: 10.1007/s10616-004-2978-2.
Characterization of genetic disorders in humans and animal models requires identification of chromosomal aberrations. However, identifying fine deletions or insertion in metaphase chromosomes has been always a challenge due to limitations of resolution. In this study we developed a rapid method for chromosome elongation using two different intercalating agents: ethidium bromide and 5-bromo-2'-deoxyuridine (BrdU), together with a short-term mitotic block using colcemid. About 70% of the chromosomes from cells that underwent this elongation procedure reached three times longer than those prepared from control cells. FISH experiments using elongated chromosomes revealed a duplicated region of chromosome 11 that was not visible in cells prepared with conventional methods.
人类和动物模型中的遗传疾病特征需要鉴定染色体异常。然而,由于分辨率的限制,鉴定中期染色体的细微缺失或插入一直是一个挑战。在这项研究中,我们开发了一种使用两种不同嵌入剂(溴化乙锭和 5-溴-2'-脱氧尿苷(BrdU))和短期秋水仙素阻断有丝分裂来使染色体伸长的快速方法。大约 70%经过这种伸长程序的细胞的染色体比用对照细胞制备的染色体长三倍。使用伸长染色体的 FISH 实验显示出 11 号染色体的一个重复区域,而在使用常规方法制备的细胞中则无法看到。