Raap AK, Florijn RJ, Blonden LAJ, Wiegant J, Vaandrager JW, Vrolijk H, Tanke HJ
Department of Cytochemistry and Cytometry, Leiden University, Wassenaarseweg 72, Leiden, NL 2333 AL, The Netherlands
Methods. 1996 Feb;9(1):67-73. doi: 10.1006/meth.1996.0009.
Fluorescence in situ hybridization (FISH) applied to metaphase chromosomes provides a mapping resolution of 1 to 3 Mb. FISH applied to interphase nuclei has a resolution of 50 kb and ranges 1-2 Mb. This better resolution is attributed to the higher degree of chromatin decondensation. Here, we describe FISH applied to naked DNA fibers (fiber FISH) and show that with such fully decondensed chromatin a resolution range of at least 1-400 kb can be obtained. Furthermore, we show that DNA fiber FISH provides a mapping tool that is highly supplementary to restriction mapping, because it permits very accurate gap and overlap sizing. Also, DNA fiber FISH provides the means to generate "color bar codes" for disease regions, which can be used to inspect patient DNAs for suspected gene rearrangements.
应用于中期染色体的荧光原位杂交（FISH）提供了1至3兆碱基的图谱分辨率。应用于间期细胞核的FISH分辨率为50千碱基，范围为1至2兆碱基。这种更高的分辨率归因于染色质更高程度的解聚。在这里，我们描述了应用于裸DNA纤维的FISH（纤维FISH），并表明对于这种完全解聚的染色质，可以获得至少1至400千碱基的分辨率范围。此外，我们表明DNA纤维FISH提供了一种与限制性图谱高度互补的图谱绘制工具，因为它允许非常精确地确定缺口和重叠的大小。而且，DNA纤维FISH提供了为疾病区域生成“彩色条形码”的方法，可用于检查疑似基因重排的患者DNA。
