摘要
目的为寻找一种高效、简便、实用的石蜡组织中DNA提取方法，本研究比较了三种不同的DNA提取方法对DNA质量的影响。方法选取淋巴相关组织的蜡块21例。三种DNA提取方法分别是简单消化煮沸法，酚／氯仿提纯法和试剂盒法。通过提取DNA的浓度和纯度、PCR扩增保守序列pglobin等方法，比较三种DNA提取方法的优劣。结果试剂盒法提取样品OD260／OD280平均比值位于1．6～1．8之间；提纯法位于1．3—1．5之间；简单消化煮沸法位于1，2—1．3之间。三种方法中，试剂盒法提取的DNA最为稳定。以3种DNA提取方法所获取DNA为模板，PCR扩增阳性率无统计学差异。结论从实用、快速、经济的角度综合分析，简单消化煮沸法简单快速，需要的组织样品少，是一个值得推荐的石蜡组织DNA提取方法。
Objectlve To seek an effective, simple and practical DNA extraction method from paraffin embedded lymphoid tissue, three chosen methods were employed in our study. Methods 21 blocks of lymphoid tissue were used in the test. Three DNA extraction methods respectively are simple digestion and boiling, phenol/chloroform pur/fication, and commercial Kit methods.Results By comparison,the OD260/OD280 average ratio is about 1.6 - 1.8 by Kit method, 1.3 - 1.5 by extraction method, 1.2- 1.3 by simple boiling method. Of the three DNA extraction methods, the Kit method is the most DNA stable one. But there is no significance in the house keeping gene-β-globin amplification among three methods. Conclusion Simply boiling DNA extraction method is simpler,quicker,more effective and much cheaper than the phenol/chloroform purification or commercial kit methods.
出处
《中国实验诊断学》
2006年第4期351-355,共5页
Chinese Journal of Laboratory Diagnosis
基金
广东省社会发展攻关项目(B30101)
广州市科技计划项目(Z3-E4061)资助
