... here ). Lentiviral plasmids can recombine between their LTRs resulting in a smaller plasmid containing...amplification of CRISPR pooled plasmid libraries in Escherichia coli cells. Protocols...starting point for CRISPR libraries. This protocol allows the amplification of a pooled-plasmid library in Escherichia...differences, and plasmid recombination can all impact the representation of individual plasmids in the pooled... Protocols CRISPR Library Amplification...Amplification CRISPR Library Amplification You may also like... Pooled libraries Guide to Using Pooled Libraries...Libraries All CRISPR Pooled Libraries Molecular Biology Reference How do I process my Addgene pooled library...

...strain Watch the Video! CRISPR Library Amplification Amplify CRISPR pooled-plasmid libraries Diagnostic ...complicated procedures. Plasmid Cloning Protocols for constructing and analyzing your plasmids. Virus Protocols...Transfection Introduce plasmid DNA to mammalian cells Lentivirus Production Produce lentivirus with a polyethyenimine... that can be used for basic molecular biology, plasmid cloning, and titering and testing your viral preparations...Bacterial Glycerol Stocks Store bacterial strains or plasmids for long term use Watch the Video! DNA Purification...spectrophotometer Restriction Digests Cut and analyze plasmid DNA with restriction enzymes Agarose Gel Electrophoresis...Diagnostic Restriction Digest Verify a plasmid using restriction enzymes Watch the Video! Polymerase Chain Reaction...

...single cells for 13 days. lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described... Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature...cells that have taken up the lentiviral transgene. Note that not all lentiviral vectors deliver antibiotic...Thaw the lentiviral aliquot on ice prior to use. Prepare a range of dilutions of the lentivirus in DMEM.... This is because lentiviral integration is random. Given the MOI of the lentivirus used, cells may also...an integrated lentiviral vector. Protocols... Protocols Generating Stable Cell Lines with Lentivirus Generating Stable...

...a loading control. 1 lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described... Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature... pool of stable cells. Transducing cells with lentivirus results in a heterogenous polyclonal population...protocol for generating stable cell lines with lentivirus . Day 0: (optional) Seed cells for the generation...protocol for generating stable cell lines with lentivirus . Pro-Tip Because the polyclonal cell pool is...expressing Cas9. A549 cells were transduced with lentiCas9-Blast 1 and then selected with 1 μg/mL blasticidin...lines generated from A549 cells transduced with lentiCas9-Blast 1 . A549 cells were transduced (MOI = 37...