Clone Assay:Article Title: A mobile CRISPRi collection enables genetic interaction studies for the essential genes of Escherichia coli.
Article Snippet: .. Method details Plasmids and construction of CRISPRi collection sgRNAs targeting different essential genes in E. coli were cloned into the conjugative CRISPRi plasmid pFD152 using a previously described single step Golden Gate assembly protocol. pFD152 was a gift from David Bikard (Addgene plasmid # 125546). sgRNAs used in this study were designed using the publicly available CRISPRbact tool (https://gitlab.pasteur.fr/dbikard/crisprbact ), wherein, an sgRNA with the highest predicted on-target activity score and the least off-target homology was selected for each gene. .. Oligonucleotides for cloning sgRNAs were ordered from Sigma Aldrich, and specific sequences of oligonucleotides and sequences of sgRNAs can be found in Table S1.
Article Title: A mobile CRISPRi collection enables genetic interaction studies for the essential genes of Escherichia coli
Article Snippet: .. sgRNAs targeting different essential genes in E. coli were cloned into the conjugative CRISPRi plasmid pFD152 using a previously described single step Golden Gate assembly protocol. pFD152 was a gift from David Bikard (Addgene plasmid # 125546). sgRNAs used in this study were designed using the publicly available CRISPRbact tool ( https://gitlab.pasteur.fr/dbikard/crisprbact ), wherein, an sgRNA with the highest predicted on-target activity score and the least off-target homology was selected for each gene. .. Oligonucleotides for cloning sgRNAs were ordered from Sigma Aldrich, and specific sequences of oligonucleotides and sequences of sgRNAs can be found in .
Article Title: A mobile CRISPRi collection enables genetic interaction studies for the essential genes of Escherichia coli
Article Snippet: .. Plasmids and construction of CRISPRi collection sgRNAs targeting different essential genes in E. coli were cloned into the conjugative CRISPRi plasmid pFD152 using a previously described single step Golden Gate assembly protocol.44 pFD152 was a gift from David Bikard (Addgene plasmid # 125546). sgRNAs used in this study were designed using the publicly available CRISPRbact tool (https://gitlab.pasteur.fr/dbikard/crisprbact), wherein, an sgRNA with the highest predicted on-target activity score and the least off-target homology was selected for each gene. .. Oligonucleotides for cloning sgRNAs were ordered from Sigma Aldrich, and specific sequences of oligonucleotides and sequences of sgRNAs can be found in Table S1.
Plasmid Preparation:Article Title: A mobile CRISPRi collection enables genetic interaction studies for the essential genes of Escherichia coli.
Article Snippet: .. Method details Plasmids and construction of CRISPRi collection sgRNAs targeting different essential genes in E. coli were cloned into the conjugative CRISPRi plasmid pFD152 using a previously described single step Golden Gate assembly protocol. pFD152 was a gift from David Bikard (Addgene plasmid # 125546). sgRNAs used in this study were designed using the publicly available CRISPRbact tool (https://gitlab.pasteur.fr/dbikard/crisprbact ), wherein, an sgRNA with the highest predicted on-target activity score and the least off-target homology was selected for each gene. .. Oligonucleotides for cloning sgRNAs were ordered from Sigma Aldrich, and specific sequences of oligonucleotides and sequences of sgRNAs can be found in Table S1.
Article Title: A mobile CRISPRi collection enables genetic interaction studies for the essential genes of Escherichia coli
Article Snippet: .. sgRNAs targeting different essential genes in E. coli were cloned into the conjugative CRISPRi plasmid pFD152 using a previously described single step Golden Gate assembly protocol. pFD152 was a gift from David Bikard (Addgene plasmid # 125546). sgRNAs used in this study were designed using the publicly available CRISPRbact tool ( https://gitlab.pasteur.fr/dbikard/crisprbact ), wherein, an sgRNA with the highest predicted on-target activity score and the least off-target homology was selected for each gene. .. Oligonucleotides for cloning sgRNAs were ordered from Sigma Aldrich, and specific sequences of oligonucleotides and sequences of sgRNAs can be found in .
Article Title: A mobile CRISPRi collection enables genetic interaction studies for the essential genes of Escherichia coli
Article Snippet: .. Plasmids and construction of CRISPRi collection sgRNAs targeting different essential genes in E. coli were cloned into the conjugative CRISPRi plasmid pFD152 using a previously described single step Golden Gate assembly protocol.44 pFD152 was a gift from David Bikard (Addgene plasmid # 125546). sgRNAs used in this study were designed using the publicly available CRISPRbact tool (https://gitlab.pasteur.fr/dbikard/crisprbact), wherein, an sgRNA with the highest predicted on-target activity score and the least off-target homology was selected for each gene. .. Oligonucleotides for cloning sgRNAs were ordered from Sigma Aldrich, and specific sequences of oligonucleotides and sequences of sgRNAs can be found in Table S1.
Activity Assay:Article Title: A mobile CRISPRi collection enables genetic interaction studies for the essential genes of Escherichia coli.
Article Snippet: .. Method details Plasmids and construction of CRISPRi collection sgRNAs targeting different essential genes in E. coli were cloned into the conjugative CRISPRi plasmid pFD152 using a previously described single step Golden Gate assembly protocol. pFD152 was a gift from David Bikard (Addgene plasmid # 125546). sgRNAs used in this study were designed using the publicly available CRISPRbact tool (https://gitlab.pasteur.fr/dbikard/crisprbact ), wherein, an sgRNA with the highest predicted on-target activity score and the least off-target homology was selected for each gene. .. Oligonucleotides for cloning sgRNAs were ordered from Sigma Aldrich, and specific sequences of oligonucleotides and sequences of sgRNAs can be found in Table S1.
Article Title: A mobile CRISPRi collection enables genetic interaction studies for the essential genes of Escherichia coli
Article Snippet: .. sgRNAs targeting different essential genes in E. coli were cloned into the conjugative CRISPRi plasmid pFD152 using a previously described single step Golden Gate assembly protocol. pFD152 was a gift from David Bikard (Addgene plasmid # 125546). sgRNAs used in this study were designed using the publicly available CRISPRbact tool ( https://gitlab.pasteur.fr/dbikard/crisprbact ), wherein, an sgRNA with the highest predicted on-target activity score and the least off-target homology was selected for each gene. .. Oligonucleotides for cloning sgRNAs were ordered from Sigma Aldrich, and specific sequences of oligonucleotides and sequences of sgRNAs can be found in .
Article Title: A mobile CRISPRi collection enables genetic interaction studies for the essential genes of Escherichia coli
Article Snippet: .. Plasmids and construction of CRISPRi collection sgRNAs targeting different essential genes in E. coli were cloned into the conjugative CRISPRi plasmid pFD152 using a previously described single step Golden Gate assembly protocol.44 pFD152 was a gift from David Bikard (Addgene plasmid # 125546). sgRNAs used in this study were designed using the publicly available CRISPRbact tool (https://gitlab.pasteur.fr/dbikard/crisprbact), wherein, an sgRNA with the highest predicted on-target activity score and the least off-target homology was selected for each gene. .. Oligonucleotides for cloning sgRNAs were ordered from Sigma Aldrich, and specific sequences of oligonucleotides and sequences of sgRNAs can be found in Table S1.
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