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Application of Making Mutants by the Method of Direct Digestion DNA in Liquified Low Melting Agarose Gel

Yuan Jing; Chen Hongjung; Lobel Peter(Department of Environmental Health, School of Public Health, Tongji Medical University,Wuhan 430030)(Center for Advanced Biotechnology and Medicine, Piscataway, USA 08854)  
In this paper,the method of making mutants by multiple polymerase chain reaction (PCR) combined with direct digestion DNA in liquified low melting agarose gel was reported. We have made 8 mutants by the method described here. It was confirmed that the positive rate of recombination DNA was 100 percent. The slice of interesting DNA was taken out from the low melting agarose gel and was completely melted at 68℃. Then the melted slice was diluted with 5 volumes of distilled deionized water. The mixture could be then directly used for digesting DNA with appropriate enzyme. The suitable percentage of the agarose gel in mixture should be less then 1%. As a result, the shortcoming was overcome that the low yield of interesting DNA after purification can impossibly be used for further experiment.
【CateGory Index】: R394
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