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Sigma Aldrich Fine Chemicals Biosciences Cas9 Lambda Red Homologous1EA
Supplier: Sigma Aldrich Fine Chemicals Biosciences CAS9BAC1P1EA
Recent publications using CRISPR/Cas9-mediated recombineering in E. coli tout editing efficiencies near 100% making CRISPR/Cas9-mediated recombineering the most powerful bacterial genome engineering method to date. In addition Cas9-mediated recombineering overcomes the dependence on a second recombination step avoids the creation of destabilizing scar sites can be used in multiplexing and is less time-consuming than previous protocols.Here we present a novel dual-vector CRISPR/Cas-mediated»-Red system for improved recombineering in E. coli. Our system is shown to facilitate homology-directed repair of DSBs created by Cas9 endonuclease enabling genetic alterations through chromosomal integration of a donor DNA.This plasmid is to be used in combination with a custom gRNA (CRISPRBACD) which can be designed and ordered through our Custom gRNA Design Tool. The donor can be either ssDNA or dsDNA with homology arms of 45-59 or 150-500 nucleotides respectively. Protocols for donor desi
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