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URL: http://www.addgene.org/69763
Proper Citation: RRID:Addgene_69763
Insert Name: 77,406 unique sgRNAs covering 20,121 genes in the human genome
Organism: Homo sapiens
Bacterial Resistance: Ampicillin
Defining Citation: PMID:26190106
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
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/
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Source: Addgene
