摘要
目的比较加热去蛋白法、碘化钠法、TT溶血法、氯化胺法四种方法提取人微量全血基因组DNA的纯度及总量的差异。方法收集静脉全血0.5ml,共356人份,分别采用上述四种方法提取基因组DNA,用紫外分光光度仪及琼脂糖凝胶电泳检测其纯度和总量,计算采用x±s,结果经统计学t检验处理。结果四种方法提取的基因组DNA纯度用OD260/OD280表示分别为:加热法1.569±0.158;碘化钠法1.517±0.093;TT溶血法1.789±0.161;氯化胺法1.815±0.051。四种方法提取的基因组DNA总量分别为:加热法(9.34±3.59)μg;碘化钠法(7.82±4.23)μg;TT溶血法(20.98±3.56)μg;氯化胺法(22.07±4.53)μg;加热法与碘化钠法比较,TT溶血法与氯化胺法比较,P>0.05,无显著性差异;TT溶血法、氯化胺法分别与加热法、碘化钠法比较,P<0.05,有显著性差异。结论从提取的基因组DNA纯度和总量来比较,氯化胺法与TT溶血法是四种提取人微量全血基因组DNA方法中较好的两种。
Objective: To compare the different purity and yield of human genomic DNA extracted from microsamples of whole blood by the methods of pyrogenation, sodium iodide, TT hemolysis and ammonium chloride. Methods: 356 samples of 0.5ml peripheral venous blood were collected, and genomic DNA was extracted from them by the four methods ahead respectively. Then the purity and yield were measured by ultraviolet photometer and aganose gel electrophoresis.The results were described as x±s and analysed with t-test. Results: DNA purity was indicated by OD260/OD280: method of pyrogenation (1.569±0.158); method of sodium iodide (1.517±0.093); method of TT hemolysis (1.789±0.161); method of ammonium chloride (1.815±0.051). There was no significant discrepancy when method of pyrogenation was compared with method of sodium iodide or method of TT hemolysis with method of ammonium chloride (P>0.05), significant discrepancy was found when method of TT hemolysis or ammonium chloride was compared with methods of pyrogenation or sodium iodide respectively(P<0.05). Conclusions: According to the purity and yield of genomic DNA, methods of ammonium chloride and TT hemolysis are the best two method of four for extracting DNA from microsamples of whole blood.
出处
《中国医学工程》
2004年第5期43-45,共3页
China Medical Engineering
关键词
微量全血
提取方法
加热去蛋白法
碘化钠法
TT溶血法
基因组DNA
genomic DNA/extraction
method of pyrogenation
method of sodium iodide
method of TT hemolysis
method of ammonium chloride.
