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Subcloning Plus Insertion (SPI) – A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
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生物学
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JoVE Journal 生物学
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

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09:02 min

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January 08, 2015

DOI:

10.3791/52155-v

DOI:
10.3791/52155-v

•

09:02 min
January 08, 2015

•

1 Views
, , , ,
1Department of Biochemistry,University of Leicester, 2Center for Fisheries, Environment and Aquaculture Sciences, 3ES Cell Facility, Centre for Core Biotechnology Services,University of Leicester

概要

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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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Cite this Article

Reddy, T. R., Kelsall, E. J., Fevat, L. M., Munson, S. E., Cowley, S. M. Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors. J. Vis. Exp. (95), e52155, doi:10.3791/52155 (2015).

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