全血基因组DNA的提取和纯化方法比较

Comparative Study of Whole Blood Genomic DNA Purification Method

目的 探讨全血基因组DNA的快速纯化方法。方法 分别采用表面活性剂裂解法、传统酚抽提法、加热法分别制备DNA ,分析各种方法提取DNA的产量、纯度 ,以及方法本身的优缺点、费用等 ,同时我们进行RAPD标记实验以比较纯化的DNA在PCR上应用的效果。结果 表面活性剂裂解法简单快速 ,其制备的DNA产率、纯度明显高于传统酚抽提法 ,但其完整性不如后者。加热法快速制备的全血基因组DNA作为模板虽然纯度差 ,但仍可用于PCR ,并得到满意的检测结果。同一份标本采用不同方法提取的DNA作为模板进行的RAPD标记实验 ,其多重PCR产物进行分离产生的DNA图谱基本一致。结论 3种全血DNA纯化方法均可满足不同的实验要求。

Objective To study a quick method of whole blood genomic DNA purificati on. Methods This article introduced three common DNA purification methods from fr esh whole blood,analyzed their quantity,purity,cost and advantage and disadvant age. At the same time,their application in PCR based on RARD marker was assesse d. Result Surfacttant lysis method is simpler and quicker.The quantity purity of t he DNA extracted was obviously better than that of the traditional phenol extr act ion method. But the integrity of the former was worse than that of the latter,Al though the purity of whole blood genomic DNA extracted by boiling method as DNA template was bad,PCR-amplification's result was still gotton. When its genomic DNA was extracted from the same blood sample by different method, respectivel y,they were used as DNA template for RAPD analysis,their DNA band patterns of m ultiple PCR products were basically in accordance with each other. Conclusion Based on different experimental objects, requirements and progress, suitable DNA purification method is chosen.