对小麦RAPD常规的扩增循环参数及琼脂糖浓度进行了试验优化。结果表明:适宜的扩增循环参数为:94℃预变性2 m in,94℃15 s→36℃15 s→72℃30 s 40个扩增循环,72℃延伸10 m in,4℃保持。用于电泳介质的琼脂糖最佳浓度为2%。时间比常规减...对小麦RAPD常规的扩增循环参数及琼脂糖浓度进行了试验优化。结果表明:适宜的扩增循环参数为:94℃预变性2 m in,94℃15 s→36℃15 s→72℃30 s 40个扩增循环,72℃延伸10 m in,4℃保持。用于电泳介质的琼脂糖最佳浓度为2%。时间比常规减少一半。展开更多
Objective： To establish a method of genomic DNA extraction from whole blood using Fe3O4/Au composite particles as a carrier. Methods： Two crucial conditions （sodium chloride concentration and amount of the magnetic...Objective： To establish a method of genomic DNA extraction from whole blood using Fe3O4/Au composite particles as a carrier. Methods： Two crucial conditions （sodium chloride concentration and amount of the magnetic particles） were optimized and 8 different human whole blood samples were used to purify genomic DNA under the optimal condition. Then agarose gel electrophoresis and polymerase cbain reaction （PCR） were performed. Results： The optimal binding condition was 1.5 mol/L NaC1/10% PEG, and the optimal amount of Fe3O4/Au composite particles was 600μg. The yields of the genomic DNA from 100μl of different whole blood samples were 2-5 μg, and the ratio of A260/A280 was in the range of 1.70-1.90. The size of genomic DNA was about 23 kb and the PCR was valid. Conclusion： The purification system using Fe3O4/Au composite microparticles has advantages in high yield, high purity, ease of operating, time saving and avoiding centrifugation. The purified sample was found to function satisfactorily in PCR amplification.展开更多
基金Supported by the National High Technology Research and Development Program of China (2006AA020705)
文摘Objective： To establish a method of genomic DNA extraction from whole blood using Fe3O4/Au composite particles as a carrier. Methods： Two crucial conditions （sodium chloride concentration and amount of the magnetic particles） were optimized and 8 different human whole blood samples were used to purify genomic DNA under the optimal condition. Then agarose gel electrophoresis and polymerase cbain reaction （PCR） were performed. Results： The optimal binding condition was 1.5 mol/L NaC1/10% PEG, and the optimal amount of Fe3O4/Au composite particles was 600μg. The yields of the genomic DNA from 100μl of different whole blood samples were 2-5 μg, and the ratio of A260/A280 was in the range of 1.70-1.90. The size of genomic DNA was about 23 kb and the PCR was valid. Conclusion： The purification system using Fe3O4/Au composite microparticles has advantages in high yield, high purity, ease of operating, time saving and avoiding centrifugation. The purified sample was found to function satisfactorily in PCR amplification.
