从福尔马林固定、石蜡包埋存档组织中提取DNA的方法比较
Comparison of methods for the extraction of DNA from formalin-fixed, paraffin-embedded archival tissues.
作者信息
Sengüven Burcu, Baris Emre, Oygur Tulin, Berktas Mehmet
机构信息
1. Department of Oral Pathology, Faculty of Dentistry, Gazi University, Ankara, Turkey;
2. Pharmacoeconomy and Pharmacoepidemiology Research Center (PEPIRC), Yeditepe University, Istanbul, Turkey.
出版信息
Int J Med Sci. 2014 Mar 27;11(5):494-9. doi: 10.7150/ijms.8842. eCollection 2014.
AIM
Discussing a protocol involving xylene-ethanol deparaffinization on slides followed by a kit-based extraction that allows for the extraction of high quality DNA from FFPE tissues.
METHODS
DNA was extracted from the FFPE tissues of 16 randomly selected blocks. Methods involving deparaffinization on slides or tubes, enzyme digestion overnight or for 72 hours and isolation using phenol chloroform method or a silica-based commercial kit were compared in terms of yields, concentrations and the amplifiability.
RESULTS
The highest yield of DNA was produced from the samples that were deparaffinized on slides, digested for 72 hours and isolated with a commercial kit. Samples isolated with the phenol-chloroform method produced DNA of lower purity than the samples that were purified with kit. The samples isolated with the commercial kit resulted in better PCR amplification.
CONCLUSION
Silica-based commercial kits and deparaffinized on slides should be considered for DNA extraction from FFPE.
目的
探讨一种方案，该方案包括对载玻片进行二甲苯 - 乙醇脱蜡，然后采用基于试剂盒的提取方法，以便从福尔马林固定石蜡包埋（FFPE）组织中提取高质量的DNA。
方法
从16个随机选择的组织块的FFPE组织中提取DNA。比较了在载玻片或试管上进行脱蜡、酶消化过夜或72小时以及使用苯酚氯仿法或基于硅胶的商业试剂盒进行分离等方法在产量、浓度和可扩增性方面的差异。
结果
通过在载玻片上脱蜡、消化72小时并使用商业试剂盒分离得到的样品产生的DNA产量最高。用苯酚 - 氯仿法分离的样品产生的DNA纯度低于用试剂盒纯化的样品。用商业试剂盒分离的样品导致更好的PCR扩增效果。
结论
从FFPE中提取DNA时应考虑使用基于硅胶的商业试剂盒并在载玻片上进行脱蜡处理。
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