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文献和实验Targeting Vector Construction Through Recombineering
Gene targeting in mouse embryonic stem cells is an essential, yet still very expensive and highly time-consuming, tool and method to study gene function at the organismal level or to create mouse models of human diseases. Conventional cloning
Preparing Vector DNA For Transfection/Targeting
Since most people have their own preferences with this task you are completely at your leisure to ignore this section. This procedure works for me. However, be aware that highly purified DNA is important for high rates of targeting
Purification of Gene Targeting Vector DNA for Electroporation
1. Purify plasmid from bacteria. We recommend the Qiagen EndoFree Plasmid Maxi kit for the purification of the targeting vector plasmid from bacteria. Please follow the directions in the kit. Electroporation of Qiagen purified DNA has been used
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