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

Tags

Subcloning Plus Insertion SPI Recombineering Method Rapid Construction Gene Targeting Vectors Homologous Recombination Targeting Vector Escherichia Coli Cells Phage Recombinases Long Homology Sequences Selection Markers Allelic Exchange Vector Assembly Multiplex Approach Plasmid Gap Repair Genomic Sequence Bacterial Artificial Chromosome Libraries Correct Recombinants
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