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目的 建立一种快速、简便、敏感检测人类基因组 DNA点突变的方法。方法 用人工诱发的带有单个碱基改变的 PCR片段作目的基因 ,借助高效毛细管电泳技术对 PCR片段作 SSCP分析。电泳采用中性毛细管涂层柱和 4%线性聚丙烯酰胺凝胶电泳缓冲液 ,紫外检测仪检测。结果 196 bp长的 PCR片段用毛细管电泳可在 2 5 min内分离出双链及单链峰 ,仅有一个碱基差异的两条单链 DNA也能得到较好地分离。结论 毛细管电泳技术可用于作 PCR- SSCP分析 ,具有快速、灵敏、准确、重复性好等特点 ,为筛选基因点突变提供了一种十分有效的方法
Objective To establish a rapid, simple and sensitive method for detecting point mutations in human genomic DNA. Methods The artificial PCR fragment with single base change was used as the target gene and SSCP analysis was performed on the PCR fragment by high performance capillary electrophoresis. Electrophoresis using neutral capillary column and 4% linear polyacrylamide gel electrophoresis buffer, UV detector detection. Results The double-stranded and single-stranded DNA were separated by capillary electrophoresis in a 196 bp PCR fragment. The two single-stranded DNAs with one base difference were separated well. Conclusion The capillary electrophoresis technique can be used for PCR-SSCP analysis. It is fast, sensitive, accurate and reproducible. It provides a very effective method for screening point mutations