We narrowed to 4 results for: cas9 plasmid
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TypeProtocol...A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9-Blast 1 and then selected with...Figure 2: Cas9 expression in monoclonal cell lines generated from A549 cells transduced with lentiCas9-Blast... express Cas9 and are included as a control (WT). Vinculin is a loading control. 1 lentiCas9-Blast was...additional 3 weeks under blasticidin selection. Anti-Cas9 Western blotting was performed on whole-cell extracts...was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide...
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Video Library
TypeProtocol...Storing and Handling Addgene Plasmids What to do after you receive your plasmids from Addgene Instructions...Protocol Diagnostic Restriction Digest Verify a plasmid using restriction enzymes Diagnostic Restriction...Instructions for Storing and Handling Plasmids Genomic Deletions with CRISPR Generate genomic deletions in mammalian...mammalian cells lines using CRISPR/Cas9 Genomic Deletions with CRISPR Protocol Over-Agar Antibiotic Plating...Transformations We walk you through a standard plasmid transformation protocol and offer some tips and...Center How to Deposit Your guide to depositing plasmids with Addgene Depositing Page Career Videos (Link... -
Virus Protocol - Generating Stable Cell Lines
TypeProtocol...A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9-Blast and then selected with...single cells for 13 days. lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described... -
Lentivirus Production
TypeProtocol...DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid encoding your insert of interest... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be...transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish* Packaging Plasmid ( psPAX2 ) 1.3...1.3 pmol Envelope Plasmid ( pMD2.G ) 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume...volume 500 μL *Plasmid concentrations and ratios should be optimized for each transfer plasmid. Pro-Tip Endotoxins...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ..., the plasmids should also be propagated in an endonuclease negative E. coli strain such as NEB stable...