...amplification of CRISPR pooled plasmid libraries in Escherichia coli cells.... 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...following protocol can be used as a starting point for CRISPR libraries. This protocol allows the amplification...Prewarm 3X LB Agar + Antibiotic plates at 37 °C. Prewarm 8X LB Agar + Antibiotic Bioassay plates. Prechill...Amplification is usually necessary to produce sufficient quantities of library for experimental applications. Repeated...

...Deletions with CRISPR Generate genomic deletions in mammalian cells lines using CRISPR/Cas9 Genomic Deletions...Deletions with CRISPR Protocol Over-Agar Antibiotic Plating Create plates to culture bacteria in the lab... lab Over-Agar Antibiotic Plating Protocol AAV Purification Purify adeno-associated virus from a preparation...

...bacterial strain Watch the Video! CRISPR Library Amplification Amplify CRISPR pooled-plasmid libraries Diagnostic...mammalian cells to produce antibodies Antibody Purification Purify recombinant antibodies using Protein A or ...testing your virus preparations. Antibodies Protocols for common antibody applications. Intro to the Lab...lab Watch the Video! Over-Agar Antibiotic Plating Quickly add antibiotic to a pre-poured plate Watch the...Formation Titering Assay for Lentivirus Quantify virus by counting antibiotic resistant colonies Generating Stable... the Video! Immunocytochemistry Use antibodies to detect antigens in cells...phenol-chloroform extract, and precipitate DNA DNA Quantification Measure DNA concentration with a spectrophotometer...

... additional 3 weeks under blasticidin selection. Anti-Cas9 Western blotting was performed on whole-cell... Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature...been sufficiently selected with the appropriate antibiotic so that every cell in the culture has, in theory...maintain cells in selective medium at a reduced antibiotic concentration (e.g., half the concentration that...by passing through a 40 µm cell strainer mesh. Quantitate the cell concentration in this homogenized cell...Sample Calculation First, use a hemocytometer to quantitate the homogenized cell solution. Homogenized cell...

... Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature...concentration of the antibiotic in culture, or remove the antibiotic entirely. If the antibiotic is reduced or...transfection. Some lentiviral vectors deliver mammalian antibiotic resistance (e.g., puromycin, blasticidin), which...stable cell culture after transduction. Performing antibiotic selection on transduced cells enables elimination...the transducibility of the cell line used, this antibiotic selection may be a vital step for obtaining a... Note that not all lentiviral vectors deliver antibiotic resistance. This protocol was established using..., treat target cells with a range of doses of antibiotic and determine the lowest dose that kills all ...