Senger G, Lüdecke H J, Horsthemke B, Claussen U
Institut für Humangenetik der Universität, Erlangen, Federal Republic of Germany.
Hum Genet. 1990 May;84(6):507-11. doi: 10.1007/BF00210799.
Physical dissection of metaphase chromosomes is the most straightforward approach for the isolation of DNA sequences from specific chromosome regions. However, conventional microdissection techniques are too crude and inefficient for analysis of the human genome. Here we describe a technique for the precise dissection of single bands from GTG-banded chromosomes. Cells from normal amniotic fluid cell cultures are harvested by the pipette method. Microdissection is performed on an inverted microscope (magnification 1250X) with the help of extended siliconized glass needles and an electronically controlled micromanipulator. Enzymatic amplification of the dissected DNA allows the construction of band-specific DNA libraries from as few as 20 dissected chromosome fragments.
中期染色体的物理切割是从特定染色体区域分离DNA序列的最直接方法。然而，传统的显微切割技术对于人类基因组分析来说过于粗糙且效率低下。在此，我们描述一种从GTG带型染色体精确切割单个条带的技术。通过移液器法收集来自正常羊水细胞培养物的细胞。在倒置显微镜（放大倍数1250X）上借助延长的硅化玻璃针和电子控制的显微操作器进行显微切割。对切割后的DNA进行酶促扩增，使得能够从少至20个切割后的染色体片段构建条带特异性DNA文库。
