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DNA repair assay using HR-RFP and NHEJ-GFP reporter cells

LW Lin Wang
JL Jingzheng Li
HZ Hu Zhou
WZ Weidao Zhang
JG Jing Gao
PZ Ping Zheng
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Mouse ESC lines containing NHEJ reporter and HR reporter, and the I-SceI expression plasmid, pcDNA3β-I-SceI were kindly provided by professor An-yong Xie, School of Medicine, Zhejiang University. mESC reporters were transfected with pcDNA3β-I-SceI and GFP or RFP expression plasmid using Neon transfection system. At three days post-transfection, cells were analyzed for RFP+ or GFP+ frequencies using a FACS LSRFortessa flow cytometer (BD), BD FACSDiva software (Version 8.0.2), and Flowjo software (Version 7.6). The gating strategies for HR-RFP+ cells and NHEJ-GFP+ cells were presented in Supplementary Fig. 8.

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