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目的建立直接将ＤＮＡ探针定位于显示Ｒ带染色体上的简便方法。方法正常人外周血淋巴细胞培养６７小时，同时加入Ｈｏｅｃｈｓｔ３３２５８和ＢＵｄＲ继续培养５～６小时，常规方法制片。标本浸于２×ＳＳＣ中，在距其１０ｃＭ的２０Ｗ紫外灯下７５℃照射２０分钟。此标本以生物素标记的ｃｏｓｍｉｄ和ＹＡＣ克隆以及ｐＢａｍＸ７进行原位杂交，Ａｖｉｄｉｎ－ＦＩＴＣ和Ａｎｔｉａｖｉｄｉｎ进行信号的检测与放大，ＰＩ复染。于Ｏ１ｙｍｐｕｓＢＸ６０型荧光显微镜下，通过ＷＩＢ滤光镜同时观察显示Ｒ带的染色体和杂交信号。结果在显微镜下，黄绿色杂交信号直接定位于红色并显示Ｒ带的染色体上。结论该方法可用于快速并准确地进行ＤＮＡ片段的染色体定位
Objective To establish a simple and direct method to locate the DNA probe on the R chromosome. Methods Normal human peripheral blood lymphocytes were cultured for 67 hours. At the same time, Hoechst33258 and BUdR were added to continue the culture for 5-6 hours. The specimen was immersed in 2 x SSC and irradiated at 75 ° C for 20 minutes with a 10 cM 20W UV lamp. This specimen was biotinylated with cosmid and YAC clones and pBamX7 in situ hybridization, Avidin-FITC and Antiavidin signal detection and amplification, PI counterstaining. Under the O1ympusBX60 fluorescence microscope, chromosomes and hybridization signals showing the R band were simultaneously observed by a WIB filter. Results Under the microscope, the yellow-green hybridization signal is located directly on the red and shows the chromosome of the R band. Conclusion This method can be used to rapidly and accurately locate the chromosomes of DNA fragments