Combinatorial CRISPR loss-of-function screen for deubiquitinases and genes related to the ubiquitin proteasome system in HCT116 cells
Combinatorial CRISPR loss-of-function screen for deubiquitinases and genes related to the ubiquitin proteasome system in HCT116 cells
At the specified time point, the cells were detached with 0.25% trypsin and pelletted The cells were grown in a humidified incubator at 37C and 5% CO2 and were regularly passaged at 70-80% confluence. Cell numbers were kept at >100 million cells. Genomic DNA was isolated using QIAamp DNA Blood Maxi Kit (Qiagen) as per manufacturer's instructions An amplicon containing guide and UMI sequences was amplified by PCR, also attaching adapter sequences.