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目的 探索适用于大规模基因组扫描的、真正自动化的微卫星基因分型实验方法。方法 应用多重 PCR将多个微卫星位点同时扩增 ,产物通过 Mega BACE毛细管电泳测序仪进行电泳 ,其结果由Genetic Profiler软件自动分析。观察 PCR效率不同时以及 PCR体系中盐离子对分型结果的影响。结果 通过 1次 PCR和电泳能得到 96个样品 10个位点的分型结果 ,且自动分型的结果精确、可靠。其基因分型的效率是平板电泳的两倍。结论 应用毛细管电泳和 Genetic Profiler自动分析软件进行微卫星基因组扫描效率高 ,节省了人力 ,有效地避免了人为差错 ,适用于基因作图、尤其是多基因疾病的基因定位、法医学及人类进化等研究。
Objective To explore a truly automated microsatellite genotyping assay suitable for large-scale genomic scanning. METHODS: Multiple microsatellite loci were simultaneously amplified by multiplex PCR. The products were electrophoresed on a Mega BACE capillary electrophoresis sequencer. The results were automatically analyzed by Genetic Profiler software. The effect of salt ion pair typing on the PCR system was observed at different times. Results The genotyping results of 10 loci in 96 samples were obtained by one-time PCR and electrophoresis, and the result of automatic typing was accurate and reliable. Its genotyping efficiency is twice that of plate electrophoresis. Conclusion Using capillary electrophoresis and Genetic Profiler automatic analysis software to carry out microsatellite genome scanning is efficient, saves manpower, effectively avoids human error, and is suitable for gene mapping, especially for the gene localization of multi-gene diseases, forensic medicine and human evolution .