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Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
 

Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Article DOI: 10.3791/52155-v • 09:03 min • January 8th, 2015
January 8th, 2015 •
1, 1, 2, *3, *1
1Department of Biochemistry, University of Leicester, 2Center for Fisheries, Environment and Aquaculture Sciences, 3ES Cell Facility, Centre for Core Biotechnology Services, University of Leicester
* These authors contributed equally

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Gene targeting methodologies can be used to generate transgenic mice with knockout, knock-in and tagged alleles. Here, we describe an improved method of recombineering in E. coli, that we term ‘subcloning plus insertion’, which can be used to generate custom gene targeting vectors rapidly.

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Keywords: Subcloning Recombineering Gene Targeting Vector Homologous Recombination Bacterial Artificial Chromosome Conditional Gene Knockout Knock-in Vector BAC Reporter Vector Selection Marker Multiplex Approach Plasmid Gap Repair
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