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. Author manuscript; available in PMC: 2020 Nov 3.
Published in final edited form as: J Vis Exp. 2019 Mar 31;(145):10.3791/59382. doi: 10.3791/59382

Figure 3: Schematic diagram representing the workflow for sgRNAs library design, construction and verification.

Figure 3:

This workflow is as follows. First, the sgRNA library was designed and cloned into a lentiviral CRISPR vector, and then the lentivirus was packaged with the sgRNA library lentiviral vector, psPAX2 and pMD2.G vectors in the HEK293T cells. Next, MOLM13 cells were infected with a low MOI (0.3) virus and these cells underwent puromycin selection. Then, the single clone was seeded in a 96-well plate. Finally, the sgRNA single clones integrated into a genome were identified by one-step RT-qPCR, Sanger sequence and Indel mutation detection.