To obtain pure DNA directly from some complex forest soils are still very difficulty at present,though many methods even commercial kits have been attempted.This paper reports an economic and efficient method for furt...To obtain pure DNA directly from some complex forest soils are still very difficulty at present,though many methods even commercial kits have been attempted.This paper reports an economic and efficient method for further purifying crude DNA extracted from forest soils with two steps.First,the crude DNA was dissolved using the extraction buffer,which removed the debris by chloroform-isoamyl alcohol,and then reprecipitated the DNA by isopropanol;second,the recovered DNA was further purified with silica spin column.Results show that 82-91% of the humic acids was removed by step one.The remaining humic acids could be completely effaced through the second step.The recovered DNA following this protocol was quite pure and ready for sensitive conventional PCR reactions.This is an economic,efficient,and timesaving method.Moreover,crude DNA extracted by other methods can be also further purified with this new way.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 Project of National Science Foundation of China (30870419, 40971151)the National Basic Research Program of China (2008CB418004)the Open Research Foundation of Key Laboratory for Modern Sylvicultural Technology of Zhejiang Province (200605), Zhejiang Forestry College,China
文摘To obtain pure DNA directly from some complex forest soils are still very difficulty at present,though many methods even commercial kits have been attempted.This paper reports an economic and efficient method for further purifying crude DNA extracted from forest soils with two steps.First,the crude DNA was dissolved using the extraction buffer,which removed the debris by chloroform-isoamyl alcohol,and then reprecipitated the DNA by isopropanol;second,the recovered DNA was further purified with silica spin column.Results show that 82-91% of the humic acids was removed by step one.The remaining humic acids could be completely effaced through the second step.The recovered DNA following this protocol was quite pure and ready for sensitive conventional PCR reactions.This is an economic,efficient,and timesaving method.Moreover,crude DNA extracted by other methods can be also further purified with this new way.
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.
