Lengauer C, Green E D, Cremer T
Institut für Humangenetik u. Anthropologie, Universität Heidelberg, Federal Republic of Germany.
Genomics. 1992 Jul;13(3):826-8. doi: 10.1016/0888-7543(92)90160-t.
Alu-PCR protocols were optimized for the generation of human DNA probes from yeast strains containing yeast artificial chromosomes (YACs) with human inserts between 100 and 800 kb in size. The resulting DNA probes were used in chromosome in situ suppression (CISS) hybridization experiments. Strong fluorescent signals on both chromatids indicated the localization of specific YAC clones, while two clearly distinguishable signals were observed in greater than or equal to 90% of diploid nuclei. Signal intensities were generally comparable to those observed using chromosome-specific alphoid DNA probes. This approach will facilitate the rapid mapping of YAC clones and their use in chromosome analysis at all stages of the cell cycle.
优化了Alu-PCR方案，用于从含有酵母人工染色体（YAC）的酵母菌株中生成人类DNA探针，这些YAC含有人源插入片段，大小在100至800kb之间。所得的DNA探针用于染色体原位抑制（CISS）杂交实验。两条染色单体上均出现强荧光信号，表明特定YAC克隆的定位，在大于或等于90%的二倍体细胞核中观察到两个清晰可辨的信号。信号强度通常与使用染色体特异性α卫星DNA探针时观察到的强度相当。这种方法将有助于YAC克隆的快速定位及其在细胞周期各个阶段的染色体分析中的应用。
