I am new to this concept and I need some guidance on designing the primers with restriction site, homologous region, and antibiotic resistance gene casseettee.
I am also looking for the suicide vectors that work best for the homologous recombination-based gene knockout. Any advice/suggestion from your end will be helpful for me.
Rongxia Chen
The most commonly used editing tool for gene knockout is the CRISPR/Cas9 system. You can choose the pTargetF plasmid as a vector for sgRNS and homologous arm construction. You can refer to the article "Multigene Editing in the Escherichia coli Genome via the CRISPR-Cas9 System."
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Rongxia Chen
This platform may help you. https://zlab.squarespace.com/guide-design-resources
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