I'm trying to edit Hek293T by using the base editing technique. I'm following JOUNG LAB gRNA CLONING PROTOCOL and cotransfecting with xBE3 editor (only one with SaKHHBE3), my sgRNA (in BPK 1520/ one with 2260) and GFP vector (Lipofectamin 2000). After two days, I do sorting by FACS (sorting with DAPI and GFP) and let the single clones grow in a 96 well plate until they're grown enough to be picked up in bigger wells (respectively 48- or 24-well plates). When they are grown inside of the bigger wells, I split and also collect the cells and do genomic DNA extraction, PCR and purification for sequencing. But I have never gotten the mutation I wanted in the single clones. I hope someone might give me an answer.
Im attaching my examples.
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