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Species: Other
Genetic Insert: dnaQ926 dam seqA emrR ugi AID(Mut7.3.1)
Vector Backbone Description: Backbone Marker:David Liu Lab; Backbone Size:2992; Vector Backbone:CloDF13; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:26443021
Comments: The authors strongly recommend that all MP-carrying strains be maintained in rich media supplemented by 25 mM glucose. The expression of the MP-borne mutators is regulated by the E. coli arabinose pBAD promoter, which is efficiently repressed at high concentrations of glucose. Please consult Badran and Liu, Nature Communications (2015) for more detailed strain maintenance protocols.
Proper citation: RRID:Addgene_69697 Copy
Species: Other
Genetic Insert: dnaQ926 dam seqA emrR ugi AID(Mut7.3.3)
Vector Backbone Description: Backbone Marker:David Liu Lab; Backbone Size:2992; Vector Backbone:CloDF13; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:26443021
Comments: The authors strongly recommend that all MP-carrying strains be maintained in rich media supplemented by 25 mM glucose. The expression of the MP-borne mutators is regulated by the E. coli arabinose pBAD promoter, which is efficiently repressed at high concentrations of glucose. Please consult Badran and Liu, Nature Communications (2015) for more detailed strain maintenance protocols.
Proper citation: RRID:Addgene_69696 Copy
Species: Homo sapiens
Genetic Insert: HURP
Vector Backbone Description: Backbone Marker:Arshad Desai (Addgene plasmid# 44432); Vector Backbone:pIC242; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_69748 Copy
Species: Homo sapiens
Genetic Insert: Cyclin B1
Vector Backbone Description: Vector Backbone:pIC194; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_69747 Copy
Species: Homo sapiens
Genetic Insert: Centrin 3
Vector Backbone Description: Backbone Marker:Arshad Desai (Addgene plasmid# 44432); Vector Backbone:pIC242; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_69746 Copy
Species: Homo sapiens
Genetic Insert: Centrin 1
Vector Backbone Description: Backbone Marker:Arshad Desai (Addgene plasmid# 44432); Vector Backbone:pIC242; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_69744 Copy
Species: Homo sapiens
Genetic Insert: DSN1
Vector Backbone Description: Backbone Marker:Arshad Desai (Addgene plasmid# 44432); Vector Backbone:pIC242; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23530067
Proper citation: RRID:Addgene_69754 Copy
Species: Homo sapiens
Genetic Insert: spc24
Vector Backbone Description: Backbone Marker:Arshad Desai (Addgene plasmid# 44432); Vector Backbone:pIC242; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_69752 Copy
Species: Homo sapiens
Genetic Insert: CENP-T aa 1-375
Vector Backbone Description: Vector Backbone:pJAG98 (pBABEblast); Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_69761 Copy
Species: Drosophila melanogaster
Genetic Insert: Thisbe (FGF8like-1)
Vector Backbone Description: Backbone Marker:?; Backbone Size:8900; Vector Backbone:pUAST-HA; Vector Types:Insect Expression, P element-based pUAST vector for Gal4-regulated expression in Drosophila.; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19515694
Proper citation: RRID:Addgene_69769 Copy
Species: Homo sapiens
Genetic Insert: 77,406 unique sgRNAs covering 20,121 genes in the human genome
Vector Backbone Description: Backbone Marker:Stephan Kissler lab; Backbone Size:8831; Vector Backbone:pLB (Addgene Plasmid #11619); Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26190106
Comments: This sgRNA library was synthesized as an oligo pool, with 77,406 unique sgRNAs covering 20,121 genes in the human genome (see attached supplemental file for genes and target sequences).
To ensure complete gene disruption, the target sites were chosen in a region close to the translation initiation site, and most genes were covered four times. It is noteworthy that the sgRNAs start with either A or G, because our previous study showed that the preferred transcription initiation nucleotide for the U6 promoter can be either A or G (Ma et al., 2014).
The sgRNA oligo pool was PCR amplified and cloned into the lentiviral vector, pLB. To test its quality, we PCR amplified the library and analyzed the amplicons with deep sequencing. The results showed that at least 90% of the sgRNAs were retained in the plasmid library (see attached supplemental file for sequencing read counts).
For more information about library use and screening, please see the associated publication (Ma et al. Cell Reports 2015, PMID: 26190106)
This library will be delivered in a microcentrifuge tube on blue ice. You will receive ~20µL of DNA at a concentration of 50ng/µL. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.
Before ordering, please review the steps for using pooled lentiviral CRISPR libraries at https://www.addgene.org/crispr/lentiviral/library/reference/
Proper citation: RRID:Addgene_69763 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol and Kanamycin
Defining Citation: PMID:27084942
Comments: Please note that parental strain (MC4100) is streptomycin-resistant.
Proper citation: RRID:Addgene_69771 Copy
Species: Homo sapiens
Genetic Insert: PCGF1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2817; Vector Backbone:pCR8/GW/TOPO; Vector Types:Bacterial Expression; Bacterial Resistance:Spectinomycin
Proper citation: RRID:Addgene_69770 Copy
Species: Other
Genetic Insert: dnaQ926 dam seqA polB
Vector Backbone Description: Backbone Marker:David Liu Lab; Backbone Size:2992; Vector Backbone:CloDF13; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:26443021
Comments: The authors strongly recommend that all MP-carrying strains be maintained in rich media supplemented by 25 mM glucose. The expression of the MP-borne mutators is regulated by the E. coli arabinose pBAD promoter, which is efficiently repressed at high concentrations of glucose. Please consult Badran and Liu, Nature Communications (2015) for more detailed strain maintenance protocols.
Proper citation: RRID:Addgene_69657 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:None
Defining Citation: PMID:27084942
Proper citation: RRID:Addgene_69778 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:27084942
Proper citation: RRID:Addgene_69776 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:None
Defining Citation: PMID:27084942
Proper citation: RRID:Addgene_69775 Copy
Species: Other
Genetic Insert: dnaQ926 dam mutSdN
Vector Backbone Description: Backbone Marker:David Liu Lab; Backbone Size:2992; Vector Backbone:CloDF13; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:26443021
Comments: The authors strongly recommend that all MP-carrying strains be maintained in rich media supplemented by 25 mM glucose. The expression of the MP-borne mutators is regulated by the E. coli arabinose pBAD promoter, which is efficiently repressed at high concentrations of glucose. Please consult Badran and Liu, Nature Communications (2015) for more detailed strain maintenance protocols.
Proper citation: RRID:Addgene_69653 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:None
Defining Citation: PMID:27084942
Proper citation: RRID:Addgene_69774 Copy
Species: Other
Genetic Insert: dnaQ926 dam seqA
Vector Backbone Description: Backbone Marker:David Liu Lab; Backbone Size:2992; Vector Backbone:CloDF13; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:26443021
Comments: The authors strongly recommend that all MP-carrying strains be maintained in rich media supplemented by 25 mM glucose. The expression of the MP-borne mutators is regulated by the E. coli arabinose pBAD promoter, which is efficiently repressed at high concentrations of glucose. Please consult Badran and Liu, Nature Communications (2015) for more detailed strain maintenance protocols.
Proper citation: RRID:Addgene_69652 Copy
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