Treatment of cells with alkaline borate buffer extends the capability of interphase FISH mapping.
Treatment of cells with alkaline borate buffer extends the capability of interphase FISH mapping.
复制标题
用碱性硼酸盐缓冲液处理细胞可扩展间期 FISH 作图的能力。
DOI：
10.1016/0888-7543(95)80049-r
复制
发表时间：
1995
期刊：
影响因子：
4.4
通讯作者：
Trask,BJ
中科院分区：
文献类型：
--
作者：
Yokota,H;vandenEngh,G;Mostert,M;Trask,BJ
间期荧光原位杂交(FISH)已被证明是一种在100kb到1-2Mb基因组分离范围内相互定位DNA序列的方法。在0.1Mb以下的距离，探针位置很少在间期染色质中被分辨。在0.1-1-Mb的范围内，相间染色质可被模拟为自由柔性链。两个探针之间的均方相间距离与线性DNA分子上两个探针之间的基因组间隔成正比。在1-2Mb以上，相间距离和基因组分离之间的关系突然变化并趋于平稳。我们使用了碱性-硼酸盐处理来扩展间期FISH作图的能力。我们在这里表明，碱性-硼酸盐处理增加了核直径，同源染色体上的探针之间的相间距离，以及同一染色体上的探针之间的距离。我们还表明，在经硼酸盐处理的核中，杂交位点之间的均方距离与基因组分离成正比，最高可达4Mb。因此，碱性-硼酸盐处理通过增加探针之间的绝对距离和扩大相间距离与基因组分离之间的简单关系的范围来增强间期FISH定位的能力。
Interphase fluorescence in situ hybridization (FISH) has been shown to be a means to map DNA sequences relative to each other in the 100 kb to 1–2 Mb genomic-separation range. At distances below 0.1 Mb, probe sites are infrequently resolved in interphase chromatin. In the 0.1- to 1-Mb range, interphase chromatin can be modeled as a freely flexible chain. The mean square interphase distance between two probes is proportional to the genomic separation between the probes on the linear DNA molecule. Above 1–2 Mb, the relationship between interphase distance and genomic separation changes abruptly and appears to level off. We have used alkaline-borate treatment to expand the capability of interphase FISH mapping. We show here that alkaline-borate treatment increases nuclear diameter, the interphase distance between probes on homologous chromosomes, and the distance between probes on the same chromosome. We also show that the mean square distance between hybridization sites in borate-treated nuclei is proportional to genomic separation up to 4 Mb. Thus, alkaline-borate treatment enhances the capability of interphase FISH mapping by increasing the absolute distance between probes and extending the range of the simple relationship between interphase distance and genomic separation.