石蜡标本DNA提取优化方案

Study on the improvement of DNA pre-extraction from formaldehyde-fixed and paraffinembedded tissues

目的 :探索从福尔马林固定石蜡包埋组织中提取出高纯度、高浓度DNA的优化方案,为临床DNA的常规检测提供快速、经济、实用的方法。方法 :选取2011年~2013年间于湖南省某大型医院收集的598例卵巢组织石蜡标本进行切片、脱蜡、DNA提取、PCR扩增、凝胶电泳及分析。结果 :提取的DNA中,69.4%的纯度能达到标准值,99%以上标本能扩增出内参。结论 :选择适合的切片厚度,脱蜡时间和方法,合理的消化酶浓度及时间,试剂的选取等均可改善提取DNA的质量。

Objective The aim of this study was to explore an optimized proposal of DNA extraction from formaldehydefixed and paraffin-embedded tissues with high purity and high concentration,so as to provide a rapid,economic and practical way for DNA routine detection.Methods 598 ovarian tissue paraffin specimens collected from a large hospital of Human province between 2011 and 2013,and were cut into slices for further dewaxing,DNA extraction,PCR amplification,gel electroplate and analysis.Results Among the extracted DNA samples,69.4% could reach the standard value in the purity and over 99% could amplify the β-action fragments by PCR successfully.Conclusion They are all helpful to better the quality of the extracted DNA that suitable slice thickness,time and of dewaxing,reasonable digestive enzyme concentration and digestion time,and the selection of reagents.