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《Journal of Northeast Agricultural University》 2011-04
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Integrating specific sequence into target gene using PCR method

XU Yaying,WANG Yong,LUO Hong,CHEN Dian(College of Horticulture,Northeast Agricultural University,Harbin 150030,China)  
An intron was introduced into a cre gene by one-step PCR,two-step PCR,three-step PCR amplification strategy,respectively,and using three primers and two templates,in order to achieve the purpose of genetic modification.In the study,it was found that by one-step PCR amplication strategy,target genes were amplified difficultly.By two-step PCR strategy,target genes could be found,but existed non-specific amplification.By three-step PCR strategy,a linear PCR was performed using the intron fragments from the above step as the primer,the resulting products from the second step were served as the template to amplify.Target genes could be amplified efficiently.It concluded that three-step PCR strategy was an ideal method.
【Fund】: 黑龙江省青年基金资助项目(QC07C46);; 东北农业大学博士启动基金资助项目
【CateGory Index】: Q78
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