Gruzdev A D, Zaĭniev G A, Reznik N A
Tsitologiia. 1975 Jun;17(6):718-20.
To localize functional loci on cytological maps of polytene chromosomes we propose to use 10-100 times stretched chromosomes. Three different ways of stretchening are briefly considered: the squash tissue preparation, when chromosomes are stretched by hydrodynamical forces; the treatment of isolated polytene chromosomes in 10-minus 4M EDTA OR 0.8M NaCL with subsequent change of these solution for saline when abrupt structural changes occur in chromosomes and they become morphologically homogeneous threads (Gruzdev and Belaya, 1973); and, finally, the use of microneedles of the micromanipulator. After an intense (ca. 100 times) stretchening, the autoradiography is sufficient to localize the loci within one micron length of double helical DNA molecule.
为了在多线染色体的细胞学图谱上定位功能位点,我们建议使用拉伸10到100倍的染色体。简要考虑了三种不同的拉伸方法:压片组织制备,即染色体通过流体动力拉伸;将分离的多线染色体置于10⁻⁴M乙二胺四乙酸(EDTA)或0.8M氯化钠中处理,随后当染色体发生突然的结构变化并变成形态均匀的细丝时,将这些溶液换成盐水(格鲁兹德夫和别拉亚,1973年);最后是使用显微操作器的微针。经过强烈(约100倍)拉伸后,放射自显影足以将位点定位在双螺旋DNA分子一微米长度范围内。