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Transgenic Mouse Facility

BAC Recombineering Service

Recombineering (recombination-mediated genetic engineering) is a novel and powerful method enabling fast and efficient construction of vectors for subsequent manipulation of mouse genomes. It is based on homologous recombination in E. coli using recombination proteins provided from a lambda phage. This technical breakthrough is precise and independent of the presence of restriction sites and the size of the DNA molecule to be modified. Furthermore this innovative technology is significantly less laborious than traditional DNA manipulation techniques and is therefore amendable to the higher-throughput needed for a centralized service facility. Because this technology is based on homologous recombination it is incredibly versatile and can be used to create vectors for gene knockout, conditional knockout, knockin, and modified BACs for transgenic production.

The Duke Recombineering Core is a full-service facility that begins with the design of the targeting strategy based on the needs of the investigator and finishes by providing linearized DNA ready for ES cell transfection as well as suggested Southern strategies for downstream screening of homologous recombination. **The Core has generated several mouse targeting vectors, including conditional knockouts and knockins. Protocols are being established to introduce other genetic markers (green fluorescent protein and Cre recombinase) into BACS for transgenic mouse and zebrafish production.

Current Fees are $3500.00 and up for the following services:

  • BAC Transgenic
  • Conventional knockin
  • Conventional knockout
  • Conditional knockout

If you are interested in BAC Recombineering services, please contact Gary Kucera at gary.kucera@duke.edu.
 

 
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