A recombineering pipeline to make conditional targeting constructs.

Jun Fu, Madeleine Teucher, Konstantinos Anastassiadis, William Skarnes, A Francis Stewart

Genomics, BioInnovationsZentrum, Technische Universitaet Dresden, Dresden, Germany.

Methods in enzymology 2010

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Here we describe an optimized recombineering protocol for the creation of conditional alleles. Recombineering is a DNA engineering methodology based on homologous recombination in Escherichia coli. It presents a number of advantages over conventional engineering methods, including the ability to place target sites for site-specific recombinases at exactly chosen positions. Consequently, complicated designs, such as conditional alleles, can be rapidly achieved. We have applied these advantages to establish a pipeline method for the fluent generation of conditional alleles which employs Cre, Flp, and Dre recombinase target sites. 2010 Elsevier Inc. All rights reserved.

Citation

Jun Fu, Madeleine Teucher, Konstantinos Anastassiadis, William Skarnes, A Francis Stewart. A recombineering pipeline to make conditional targeting constructs. Methods in enzymology. 2010;477:125-44