Haaf T, Ward D C
Department of Genetics, Yale University School of Medicine, New Haven, CT 06510.
Hum Mol Genet. 1994 Apr;3(4):629-33. doi: 10.1093/hmg/3.4.629.
The general usefulness of fluorescence in situ hybridization (FISH) for the physical mapping of the human genome is greatly enhanced by improved DNA resolution. Several techniques have been described for decondensing or stretching interphase chromatin in a linear fashion, allowing long-range FISH mapping in finer detail. Highly extended linear chromatin can be hybridized over physical distances of at least several megabases, possibly over the whole length of a chromosome. By multi-color FISH, we have determined the order and overlaps of YACs from a 5q34-35 contig. Cosmids can be localized within larger YAC clones. Extended chromatin mapping can be applied as an adjunct ordering technique in genome studies.
通过提高DNA分辨率,荧光原位杂交(FISH)在人类基因组物理图谱绘制中的总体实用性得到了极大提升。已经描述了几种以线性方式使间期染色质解聚或伸展的技术,从而能够更详细地进行长距离FISH图谱绘制。高度伸展的线性染色质可以在至少几个兆碱基的物理距离上进行杂交,甚至可能覆盖整个染色体长度。通过多色FISH,我们确定了来自5q34 - 35重叠群的酵母人工染色体(YAC)的顺序和重叠情况。黏粒可以定位在更大的YAC克隆内。伸展染色质图谱绘制可作为基因组研究中的一种辅助排序技术应用。