...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...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette CO 2 incubator Pipet controller Hazardous waste container Microscope Hemocytometer... by passing several times through a serological pipet or by passing through a 40 µm cell strainer mesh...

... deletions in mammalian cells lines using CRISPR/Cas9 Genomic Deletions with CRISPR Protocol Over-Agar...Mulitchannel Pipetting Technique Learn how to pipette multiple samples at once using a multichannel pipette and...and avoid inconsistent pipetting with our simple tips Bacterial Transformations We walk you through a standard...

...A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9-Blast and then selected with...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM...DMEM complete + 10 µg/mL polybrene. Mix well by pipetting or inverting the tube. Aliquot 1 mL of cell suspension...formed by expansion of single cells for 13 days. lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid...

...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Inverted fluorescence...cells into 14 mL of DMEM complete. Mix well by pipetting or inverting. Aliquot 2 mL of cell suspension ...transduced with a range of dilutions of pHAGE-TO-dCas9-3XmCherry . 72 h post transduction, cells were assayed...

...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container pH meter Stir...per 10 cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would...out of the previously seeded 10 cm plate. Slowly pipette the transfection mix onto the 10 cm plate, being...

...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM...complete containing 10 μg/mL polybrene. Mix well by pipetting or inverting. Aliquot 1.35 mL of the cell suspension...serial dilutions of the lentiviral vector pHAGE EF1α dCas9-KRAB . The following day, cells were treated with...