...Addgene's Business Development Manager, discusses his transition from the Scientist Team to Business Team,...CRISPR Generate genomic deletions in mammalian cells lines using CRISPR/Cas9 Genomic Deletions with CRISPR ...

...the RNAi consortium for targeting human and mouse genes. Virus Name Description Link to Video General Transfection...antibiotic resistant colonies Generating Stable Cell Lines with Lentivirus Genomically integrate your DNA sequence...Population by Limiting Dilution Generate monoclonal cell lines from a polyclonal pool of stable cells AAV Production...

... but can be adapted to a variety of target cell lines and selection markers. Note, because this protocol...Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment...transduction efficiency. Titer will vary between cell lines. It is not recommended to subject lentiviral supernatants...

... can be modified for alternative packaging cell lines or transfection reagents. Once produced, lentivirus...Heat-inactivated FBS 1x PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment... cells in a T75 flask in 15 mL DMEM Complete. Wednesday: Plate 1×10 6 cells in a T75 flask in 15 mL DMEM...

...migrate through the gel more quickly than longer ones. Thus, you can determine the approximate length ...you can infer the size of the DNA in your sample lanes. For more details on doing diagnostic digests and...A few simple ways to increase the resolution (crispness) of your DNA bands include: a) running the gel...

... to the appropriate well. Pro-Tip Leaving clear lanes between samples will make quantifying the gel easier...Gels . Select Select Next Lane . Repeat for all lanes. Plot the area under the curves for the protein ...Figure 3 Select Analyze . Select Gels . Select Plot Lanes . One graph per lane will appear with peaks representing...

...Reagents Adherent HEK293T cells (ideally AAV-293T clones) Pro-Tip While adherent, these cells are very loosely...Fisher, 12605010) 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment... + 60 mL of 5 M Sodium Chloride + 4 mL of 1 M Magnesium Chloride Close the bottle and mix by inverting...

...short region of DNA: guide RNAs targeting different genes in an organism's genome, for example. Amplification...altering the composition of the library. Bottlenecks, fitness differences, and plasmid recombination can all ...being lost from the pool and/or increasing the skewness of the pool. 8 electroporation cuvettes (BioRad...

...These guidelines summarize the proper personal protective equipment (PPE) for BSL-1 and BSL-2 labs....Gloves Eye protection and face shields, as needed Guidelines Your lab coat should be buttoned to protect your...

...This approach can be adapted for different cell lines and different transfection reagents. Workflow Timeline...cells in a 75 cm 2 flask in a volume of 15 mL. Wednesday: Plate 1x10 6 cells in a T75 flask in a volume...