REPETITIVE SEQUENCE-DEPLETE LIBRARIES OF DNA PROBES

对于拟议的I期研究，我们计划采用新设计的减法策略从微解剖DNA中去除重复序列，并为微解剖手臂探针生成新的重复序列耗尽文库。荧光原位杂交（FISH）技术于1980 - 2015年发展起来，彻底改变了细胞遗传学的研究。人类全染色体涂绘探针、染色体臂涂绘探针和染色体带特异性涂绘探针在单色和多色FISH、光谱核型和间期FISH中的应用，极大地促进了染色体异常在癌症和遗传性疾病研究中的应用。我们的新方法将简化研究人员在FISH中使用DNA探针的协议。它将满足先进的FISH技术的要求。在第一阶段，我们将为染色体臂5p， 9q， 12p和15q以及染色体末端带12qter和18qter生成重复序列缺失DNA绘制探针和文库。在第二阶段，我们将为所有人类染色体、臂和末端带生成重复序列耗尽DNA绘制探针和文库。建议的商业应用：我们的新方法将大大降低DNA探针的生产成本。简化了FISH的程序，节省了科研人员的时间和劳动，同时又满足了先进技术的要求。这种新方法产生的DNA探针将在世界范围内得到广泛应用。

For the proposed Phase I research, we plan to apply a newly designed subtraction strategy to remove repetitive sequences from microdissected DNA, and to generate new repetitive sequence-deplete libraries for microdissected arm probes. fluorescence in situ hybridization (FISH) technique developed in 198015 revolutionized cytogenetics study. Application of human whole chromosome painting probes, chromosome arm painting probes and chromosome band-specific painting probes in single and multi-color FISH, spectrum karyotyping and interphase FISH has greatly facilitated chromosome abnormality studies in both cancer and hereditary disease research. Our new method will simplify the protocol for research scientists using DNA probes in FISH. it will meet the demands of advanced FISH techniques. During Phase I, we will generate repetitive sequence-deplete DNA painting probes and libraries for chromosome arms 5p, 9q, 12p and 15q and chromosome terminal bands 12qter and 18qter. During Phase II, we will generate repetitive sequence-deplete DNA painting probes and library for all human chromosomes, arms and terminal bands. PROPOSED COMMERCIAL APPLICATIONS: Our new method will greatly reduce the costs of DNA probes production. It will simplify protocol for FISH, thus saving time and labor for research scientist, and yet meets the demand of advanced techniques. The DNA probes generated by this new method will be utilized world wide.